EP1983056A1 - Method for transforming monocotyledons - Google Patents
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- EP1983056A1 EP1983056A1 EP08009937A EP08009937A EP1983056A1 EP 1983056 A1 EP1983056 A1 EP 1983056A1 EP 08009937 A EP08009937 A EP 08009937A EP 08009937 A EP08009937 A EP 08009937A EP 1983056 A1 EP1983056 A1 EP 1983056A1
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
- C12N15/8202—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by biological means, e.g. cell mediated or natural vector
- C12N15/8205—Agrobacterium mediated transformation
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- the present invention relates to a method for transforming monocotyledons.
- Conventional methods for transforming monocotyledons include electroporation method, polyethylene glycol method (PEG method), particle gun method and so on.
- the electroporation method is a method in which protoplasts and the desired DNA are mixed, and holes are formed in the cell membranes by electric pulse so as to introduce the DNA into the cells, thereby transforming the cells.
- This method currently has the highest reproducibility of the conventional methods and various genes have been introduced into monocotyledons, especially into rice plants by this method ( Toriyama K. et al., 1988; Bio/Technol. 6:1072-1074 , Shimamoto K. et al., 1989; Nature 338:274-276 , Rhodes C.A. et al., 1989; Science 240:204-207 ).
- this method has the problems that 1) it can be applied only to the plant species for which the system for regenerating plants from protoplasts has been established, 2) since it takes several months to regenerate plants from the protoplasts, a long time is required to obtain transformants, and that 3) since the culture period is long, the frequency of emergence of mutants during the culture is high accordingly, so that the probability of obtaining normal transformants is decreased.
- the PEG method is a method in which the desired gene and protoplasts are mixed and the mixture is treated with PEG, thereby introducing the gene into the protoplasts.
- This method is different from the electroporation method in that PEG is used instead of the electric pulse.
- the efficiency of introducing the gene is thought to be somewhat lower than the electroporation method.
- transformants were obtained by this method, this method is not widely used. Since protoplasts are used, this method has the same problems as in the electroporation method ( Zhang W. et al., 1988; Theor. Appl. Genet. 76:835-840 , Datta S.K. et al., 1990; Bio/Technol. 8:736-740 ).
- the particle gun method is a method in which the desired gene is attached to fine metal particles, and the metal particles are shot into cells or tissues at a high speed, thereby carrying out the transformation.
- transformation may be performed on any tissues. Therefore, this method is effective for transforming the plant species for which the systems for regenerating plants from protoplasts have not been established.
- the efficiency of transformation varies depending on the selection after the gene was shot. There is no data which compare the efficiency of this method with that of the electroporation method ( Gordon-Kamm W.J. et al., 1990; Plant Cell 2:603-618 , Fromm M.E. et al., 1990; Bio/Technol. 8:833-839 , Christou P. et al., 1991; Bio/Technol. 9:957-962 ).
- a method for introducing a gene using the Ti plasmid of bacteria belonging to genus Agrobacterium as a vector is widely used for transforming dicotyledons such as tobacco, petunia, rape and the like.
- dicotyledons such as tobacco, petunia, rape and the like.
- the hosts of the bacteria belonging to genus Agrobacterium are restricted to dicotyledons and that monocotyledons are not parasitized by Agrobacterium ( De Cleene M. 1976; Bot. Rev. 42:389-466 ).
- an objective of the present invention is to provide a method for transforming monocotyledons, with which the time required for obtaining regenerated plants from the time of transformation is shorter than that in the conventional methods, which can be generally applied even to the plants for which the systems for regenerating plants from protoplasts have not yet been established, and with which the preparation of the materials to be used is easy.
- the present inventors intensively studied the influences of the monocotyledonous plant tissues treated with Agrobacterium, treatment conditions of Agrobacterium, constitution of the binary vector and the like on the introduction efficiency of the genes to discover that cultured tissues of monocotyledons can be transformed by using Agrobacterium with drastically high efficiency and reproducibility and that by employing this method, the above-mentioned object can be attained, thereby completing the present invention.
- the present invention provides a method for transforming a monocotyledon comprising transforming a cultured tissue during dedifferentiation process or a dedifferentiated cultured tissue of said monocotyledon with a bacterium belonging to genus Agrobacterium containing a desired gene.
- the present invention it was first attained to introduce foreign genes to monocotyledons such as plants belonging to family Gramineae including rice, maize, wheat, barley and the like with good reproducibility.
- monocotyledons such as plants belonging to family Gramineae including rice, maize, wheat, barley and the like.
- methods for transforming monocotyledons using Agrobacterium are known, they are not established methods as mentioned above.
- Agrobacterium by inoculating Agrobacterium to cultured tissues which have not been employed in the conventional methods, genes can be very easily ' introduced.
- a cultured tissue such as a callus which can be easily prepared is used, the sample materials can be obtained more easily than the conventional method employing the apical meristem.
- Fig. 1 shows the structure of pTOK162 which is an example of the plasmid contained in Agrobacterium bacteria that may be employed in the method of the present invention, and shows the method for constructing a plasmid pTOK232 used in the examples of the present invention.
- the monocotyledons which may be transformed by the method according to the present invention are not restricted and the method according to the present invention may be applied to any monocotyledons such as rice, maize, barley, wheat, asparagus and the like.
- the cultured tissue used in the method of the present invention is a cultured tissue during dedifferentiation process or a dedifferentiated cultured tissue.
- cultured tissue during the dedifferentiation process herein means a tissue which is obtained by culturing an explant on a medium containing a plant growth regulator such as an auxin or a cytokinin, which tissue is before the stage that a callus or an adventitious embryo-like tissue is formed.
- dedifferentiated tissue herein means a callus or an adventitious embryo-like tissue obtained by culturing an explant in a medium containing a plant growth regulator such as an auxin or a cytokinin.
- the cultured tissue employed in the present invention may be originated from any part of the plant.
- cultured tissues originated from scutellum, shoot apex, root, immature embryo, pollen and anther can be employed.
- the cultured tissue employed in the present invention it is preferred to employ a cultured tissue during callus-formation process which is cultured for not less than 7 days after an explant is placed on a dedifferentiation-inducing medium, a callus or an adventitious embryo-like tissue. In the interim, it is best preferred to employ a callus or an adventive embryo-like tissue as the cultured tissue.
- Dedifferentiation-inducing media are well-known in the art. For example, a medium containing major inorganic salts and vitamins of N6 medium ( Chu C.C. 1987; Proc. Symp.
- Agrobacterium used for the transformation Agrobacterium which are employed for the transformation of dicotyledons can be employed. Many of these Agrobacterium contain a vector having a DNA region originated from the virulence region ( vir region) of Ti plasmid originated from Agrobacterium tumefaciens. The gene encoding a character which is desired to be given to the plant is inserted in this vector, or exists in a separate plasmid and inserted into the Ti plasmid in vivo by homologous recombination or the like.
- the present inventors previously developed a vector containing a DNA region originated from the virulence region ( vir region) of Ti plasmid pTiBo542 ( Jin S.
- this vector is also called "super binary vector"). Such a super binary vector may preferably be employed in the present invention.
- pTOK162 Japanese Laid-open Patent Application (Kokai) No. 4-222527 , EP-A-504,869 , U.S. Serial No. 07/854,844 ).
- the structure is shown in Fig. 1 .
- This plasmid comprises a plasmid called pTOK154 which can replicate in both Escherichia coli and in Agrobacterium tumefaciens
- pTOK154 is a plasmid containing T region, which was constructed by the method described below from a known plasmid pGA472 derived from the Ti plasmid and a known plasmid having a wide host spectrum called pVCK101), into which a Kpn I fragment (containing virB, virG and virC genes) with a size of 15.2 kb originated from the virulence region of pTiBo542 is inserted, the Kpn I fragment having been cloned.
- a kanamycin-resistant gene is inserted as a gene to be introduced into a monocotyledon.
- the gene desired to be introduced into the monocotyledon is arranged in a plasmid having the cloned DNA fragment originated from the virulence region of pTiBo542.
- each reference symbol represents the following meaning:
- the gene which is desired to be incorporated into the monocotyledon may be inserted into a restriction site in the T region of the above-described plasmid, and the desired recombinant plasmid may be selected depending on an appropriate selection marker such as drug resistance and the like which the plasmid has.
- the vector like pTOK162 shown in Fig. 1
- the desired DNA can be inserted in the T region of pTOK162 by utilizing the in vivo homologous recombination ( Herrera-Esterella L. et al, 1983; EMBO J.
- pTOK162 is preliminarily introduced into Agrobacterium tumefaciens and the plasmid pBR322 (or a similar plasmid) containing the desired gene is further introduced into Agrobacterium tumefaciens . Since pTOK162 has a region homologous with a region of pBR322, the pBR322 derivative containing the desired gene is inserted into pTOK162 by the genetic recombination via the homologous regions.
- pTOK162::pBR322 derivative the recombined pTOK162 and pBR322 is hereinafter designated "pTOK162::pBR322 derivative").
- the present inventors made a study by introducing various plasmids into Agrobacterium tumefaciens containing pTOK162, to discover that as the selection marker of the pBR322 derivative, spectinomycin-resistant gene (SP) originated from transposon Tn7 ( De Greve H.H. et al., 1981; Plasmid 6:235-248 ) is suited.
- SP spectinomycin-resistant gene
- a plasmid containing the DNA from pBR322 and SP gene is first provided, and the desired gene may be inserted into this plasmid.
- the desired gene may be inserted into this plasmid.
- the border sequences of the T region it is possible to arrange the kanamycin-resistant gene and the desired gene in separate T regions in pTOK162.
- the introduction of the desired gene can be sufficiently attained.
- both T regions may be inserted into different chromosomes, it may be possible to subsequently segregate the desired gene from the kanamycin-resistant gene.
- the gene which is desired to be introduced to monocotyledons is not restricted at all and may be any gene which can give a desired character.
- Examples of the desired gene include herbicide-resistant genes, antibiotic-resistant genes, virus coat protein genes for giving resistance to the virus, genes related to starch formation in albumen and the like, although the desired genes are not restricted thereto.
- Agrobacterium tumefaciens may preferably be employed, although not restricted.
- Agrobacterium tumefaciens Introduction of a plasmid into the bacterium belonging to genus Agrobacterium such as Agrobacterium tumefaciens can be carried out by a conventional method such as triple cross method of bacteria ( Ditta G. et al., 1980; Proc. Natl. Acad. Sci. USA 77:7347-7351 ).
- the gene which is desired to be introduced into the monocotyledon is arranged between border sequences of the T region as in the prior art, the desired gene may be arranged in the Ti plasmid or in another plasmid in the Agrobacterium .
- the transformation of the cultured tissue of a monocotyledon by the Agrobacterium may be carried out by merely contacting the cultured tissue with the Agrobacterium .
- a cell suspension of the Agrobacterium having a population density of 10 6 -10 11 cells/ml is prepared and the cultured tissue is immersed in this suspension for 3 - 10 minutes.
- the resulting culture tissue is then cultured on a solid medium for several days together with the Agrobacterium .
- transformation may be carried out by adding the Agrobacterium to the culture medium of the cultured tissue and continuously culturing the cultured tissue together with the Agrobacterium.
- the cultured tissue may be subjected to the transformation without pretreatment such as treating the cultured tissue with an enzyme or injuring the cultured tissue.
- the transformed cells or transformed tissues After subjecting the cultured tissues to the transformation, it is preferred to select the transformed cells or transformed tissues in the state during dedifferentiation process or in the dedifferentiated state. This can be attained by culturing the treated tissues on a medium containing a plant growth regulator such as an auxin or a cytokinin, which contains a selectable marker such as hygromycin and an antibiotic against the Agrobacterium , that is employed for the selection by the selection marker.
- a plant growth regulator such as an auxin or a cytokinin, which contains a selectable marker such as hygromycin and an antibiotic against the Agrobacterium , that is employed for the selection by the selection marker.
- the selected cells or the selected tissues may be subjected to regeneration culture by a known method. By this, plants acquired the desired character by the transformation can be regenerated.
- Mature seeds of rice were sterilized by being immersed in 70% ethanol for 1 minute and then in 1% sodium hypochlorite solution for 30 minutes. The seeds were then placed on 2N6 solid medium (inorganic salts and vitamins of N6 ( Chu C.C., 1978; Proc. Symp. Plant Tissue Culture, Science Press Peking, pp.43-50 ), 1g/l of casamino acid, 2 mg/l of 2,4-D, 30 g/l of sucrose, 2 g/l of Gelrite). Scutella were removed from the seeds on Day 4 from the beginning of the culture on 2N6 solid medium and used as "scutellum" samples.
- the formed calli originated from scutella were transferred to 2N6 medium and cultured therein for 4 - 7 days.
- the resulting calli were used as "scutellum callus" samples.
- Mature seeds of rice were sterilized by the above-described method and were placed on 1/2 N6 solid medium (half strength of major inorganic salts and minor salts of N6, vitamins of N6, 1 g/l of casamino acid, 20 g/l of sucrose and 2 g/l of Gelrite). From seedlings on Day 3 after germination , tissues of 2 - 3 mm length containing apex dividing tissues were cut out and used as samples.
- the calli originated from scutella were transferred to AA liquid medium (major inorganic salts of AA, amino acids of AA and vitamins of AA ( Toriyama and Hinata 1985; Plant Science 41:179-183 ), MS minor salts ( Murashige and Skoog 1962; Physiol. Plant. 15:473-497 ), 0.5 g/l of casamino acid, 1 mg/l of 2,4-D, 0.2 mg/l of kinetin, 0.1 mg/l of gibberellin and 20 g/l of sucrose) and the cells were cultured therein at 25°C in the dark under shaking of 120 rpm to obtain suspended cultured cells. The medium was replaced with fresh medium every week.
- Hygromycin resistant gene (HPT) and ⁇ -D-glucuronidase (GUS) gene were inserted in the T-DNA region of Ti plasmid to obtain the following plasmids:
- the Cla I fragment (2.5 kb) of the spectinomycin-resistant gene originated from Tn7 were treated with Klenow fragment to blunt the ends.
- the resulting fragment was inserted in Sma I site of pUC19 to obtain a plasmid pTOK107 (5.2 kb) having ampicillin-registant and spectinomycin-resistant genes.
- the obtained pTOK107 was treated with Eco RI and Hind III and the obtained 2.5 kb fragment containing the spectinomysin-resistant gene was ligated to a Eco RI - Hind III fragment (2.7 kb) of pGA482 to obtain pTOK170 (5.2 kb) containing the spectinomycin-resistant gene and has Hind III site and Hpa I site.
- a vector pIG221 in which the first intron of catalase of caster bean and GUS gene are ligated to 35S promoter was digested with Eco RI and the resultant was treated with Klenow fragment to blunt the ends.
- a Hind III linker pCAAGCTTG, code 4660P commercially available from TAKARA SHUZO.
- a fragment containing 35S promoter and intron GUS was cut out by digesting the resulting vector with Hind III, and the fragment was inserted into the Hind III site of a plasmid pGL2 (J.
- pGL2-IG 7.6 kb
- the above-mentioned plasmid pGL2 was obtained by inserting a hygromycin-resistant gene ( Gritz L. and Davis J. 1983; Gene 25: 179 - 188 ) into pDH51 ( Pietrazak et al., 1986; Nucleic Acids Research 14: 5857 - 5868 ).
- the fragment obtained by treating pTOK170 with Hpa I was ligated to a Pvu II fragment (5.2 kb) of pGL2-IG to obtain pTOK229 (10.1 kb).
- Insertion of the desired genes (hygromycin-resistant gene and intron GUS gene) into a super binary vector pTOK162 obtained by inserting virB, virC and virG genes of strongly virulent Agrobacterium tumefaciens A281 into the binary vector was carried out by homologus recombination. That is, since both vectors contain a region originated from an E. coli plasmid pBR322, in the bacterial cells selected by resistances to spectinomycin and kanamycin, only the plasmid generated by recombination of the both plasmids are contained.
- the plasmid obtained by the fact that the hygromycin-resistant gene and the intron GUS gene were inserted into the super binary vector is called pTOK232 (see Fig. 1 ).
- Strains LBA4404 and EHA101 in which T-DNA regions were deleted were used as the host bacteria.
- Strain LBA4404 has a helper plasmid PAL4404 (having a complete vir region), and is available from American Type Culture Collection (ATCC 37349).
- Strain EHA101 has a helper plasmid having the vir region originated from a strongly virulent Agrobacterium tumefaciens A281, and is available from Hood E.E. et al., 1986.
- the various binary vectors described in (2) were introduced into these two strains of Agrobacterium tumefaciens, and the strains described in the following were used for introducing the genes.
- the plasmids were introduced into the Agrobacterium strains by triple cross ( Ditta G. et al., 1980; Proc. Natl. Acad. Sci. USA 77: 7347-7351 ).
- Colonies obtained by culturing the Agrobacterium strains on AB medium ( Drlica K.A. and Kado C.I. 1974; Proc. Natl. Acad. Sci. USA 71:3677-3681 ) containing hygromycin (50 ⁇ g/ml) and kanamycin (50 ⁇ g/ml) for 3 - 10 days were collected with a platinum loop and suspended in modified AA medium (same as the composition of the above-described AA medium except that concentrations of sucrose and glucose were changed to 0.2 M and 0.2 M, respectively, and that 100 ⁇ M of acetosyringone was added, pH 5.2). The cell population was adjusted to 3 x 10 9 - 5 x 10 9 cells/ml and the suspensions were used for inoculation.
- the sample tissues were washed with sterilized water and immersed in the above-described suspensions of Agrobacterium strains, respectively, for 3 - 10 minutes. Thereafter, the shoot apex samples were placed on N6S3 solid medium (1/2 N6 major inorganic salts, N6 minor salts, N6 vitamins, Chu C.C., 1978, AA amino acids (Toriyama and Hinata 1985), 1 g/l of casamino acid, 0.2 mg/l of NAA, 1.0 mg/l of kinetin and 3 g/l of Gelrite) containing 100 ⁇ M of acetosyringone, 10 g/l of glucose and 20 g/l of sucrose.
- N6S3 solid medium 1/2 N6 major inorganic salts, N6 minor salts, N6 vitamins, Chu C.C., 1978, AA amino acids (Toriyama and Hinata 1985), 1 g/l of casamino acid, 0.2 mg/l of NAA, 1.0 mg/l of kinetin
- tissue samples such as scutellum callus samples were cultured on 2N6 solid medium containing acetosyringone, glucose and sucrose in the same concentrations as mentioned above.
- the both culture was carried out at 25°C in the dark for 2 - 5 days.
- the resulting inoculated tissues were then washed with sterilized water containing 250 mg/l of cefotaxime and then continued to be cultured on the respective solid media containing the same concentration of cefotaxime as mentioned above.
- the tissues were immersed in 0.1 M phosphate buffer (pH 6.8) containing 0.1% TRITON X-100 at 37°C for 1 hour.
- phosphate buffer containing 0.1 mM of 5-bromo-4-chloro-3-indolyl- ⁇ -D-gluconic acid and 20% methanol was added to the tissues.
- the number of blue-colored tissues were counted under a microscope and the percentages thereof based on the number of samples are described.
- Scutella cultured with the Agrobacterium strains for 3 days were cultured on 2N6 medium containing 250 mg/l of cefotaxime for 1 week. Selection of transformed cells were then carried out on 2N6 medium containing 50 mg/l of hygromycin.
- Tissues cultured with the Agrobacterium strains for 3 days were cultured on 2N6 medium containing 250 mg/l of cefotaxime for 1 week.
- Hygromycin-resistant cultured tissues were selected by culturing the cultured tissues on 2N6 medium containing 50 mg/l of hygromycin for 3 weeks (primary selection).
- the obtained resistant tissues were further cultured on N6-12 medium (N6 inorganic salts, N6 vitamins, 2 g/l of casamino acid, 0.2 mg/l of 2,4-D, 0.5 mg/l of 6BA, 5 mg/l of ABA, 30 g/l of sorbitol, 20 g/l of sucrose and 2 g/l of Gelrite) containing 50 mg/l of hygromycin for 2 - 3 weeks (secondary selection), and the calli grown on this medium were transferred to a plant regeneration medium N6S3 containing 0, 20 or 50 mg/l of hygromycin. In all of the media used after the culture with Agrobacterium strains, cefotaxime was added to 250 mg/l.
- N6-12 medium N6 inorganic salts, N6 vitamins, 2 g/l of casamino acid, 0.2 mg/l of 2,4-D, 0.5 mg/l of 6BA, 5 mg/l of ABA, 30 g/l of sorbitol, 20
- the cells cultured with the Agrobacterium strains for 5 days were cultured in 2N6 medium containing 250 mg/l of cefotaxime for 1 week, and then the selection of the transformed cells was carried out on 2N6 medium containing 50 mg/l of hygromycin.
- Seeds of the progeny of the transformants were sown in aqueous 400-fold Homai hydrate (Kumiai Kagaku Inc.) solution containing 70 mg/l of hygromycin and incubated therein at 25°C for 10 days, thereby examining the resistance to hygromycin. Twenty seeds of each plant of the progeny of the transformants were sown and cultured for about 3 weeks. From the obtained seedlings, leaves were collected and examined for the expression of GUS gene.
- the Southern blotting analysis was carried out in accordance with Molecular Cloning (Sambrook et al., 1989; Cold Spring Harbor Laboratory Press ). Two GUS positive plants, two GUS negative plants and two hygromycin-resistant plants were picked up from each of the two lines of the progeny of transformants of Tsukinohikari and were subjected to the Southern blotting analysis in the same manner as mentioned above.
- various tissues of the rice variety Tsukinohikari were treated with Agrobacterium tumefaciens EHA101 having a super-virulent vir region, into which the binary vector ( supra ) containing the hygromycin-resistant gene and the GUS gene were introduced, and then the GUS activities were examined.
- the sample tissues were shoot apices, radicles, scutella, radicle calli, scutellum calli and suspended cultured cells. In cases where the tissues were not treated with the Agrobacterium strain, no tissues exhibited GUS activity indicated by blue color.
- the binary vector pIG121Hm used in this experiment does not express GUS gene in Agrobacterium cells because the intron of caster oil plant is inserted in the promoter of the GUS gene (Nakamura et al., 1991).
- the expression of the GUS gene after the culturing with Agrobacterium is used as an index, it was confirmed that genes can be introduced into rice cells by Agrobacterium.
- shoot apex is a tissue containing the apical meristem, in order that cells expressing a hygromycin-resistance keep to grow after the treatment for introducing the gene, it is necessary that the gene be introduced into the limited apical meristem.
- the reason why no resistant tissues were obtained in spite of the fact that a number of genes were introduced into the shoot apices after the culture with the Agrobacterium strain is thought to be that the probability that the gene is introduced in the vicinity of the apical meristem is low. Further, it is easily assumed that even if a gene is introduced in the vicinity of the apical meristem so that transformed cells are obtained, the possibility that the obtained plants exhibit chimeric properties is high. From these, it is thought that the transformation method utilizing shoot apex, which was reported by Gould et al (1991) has more technical difficulties and less reproducibility than the method utilizing a dedifferentiated tissue such as callus.
- EHA101(pIG121Hm) has a helper plasmid containing the vir region of super-virulent Agrobacterium tumefaciens A281.
- LBA4404(pIG121Hm) has an ordinary vir region.
- the vir region of the helper plasmid in LBA4404(pTOK232) is ordinary, a gene which is a part of the vir region of the super-virulent Agrobacterium tumefaciens A281 is contained in the binary vector.
- This binary vector is originated from pTOK162 and made it possible to transform at a very high rate dicotyledonous species which are difficult to transform ( Saito Y. et al., 1992; Theor.
- LBA4404(pTOK232) having a part of the super-virulent vir gene in the binary vector is the best as the Agrobacterium strain used for transforming rice.
- the thus obtained resistant calli were subjected to secondary selection, and plants were regenerated from the selected resistant calli.
- a group in which hygromycin was not added to the N6S3 medium for regeneration was provided. In this group, a number of plants which did not exhibit GUS activity or which exhibited GUS activity chimerally emerged. However, in cases where hygromycin was added to the regeneration medium, the number of these plants largely decreased and the number of plants each of which exhibits GUS activity in the whole plant was increased (Table 5, Table 6 and Table 7). In cases where the tissue was not treated with Agrobacterium , no plants which exhibited resistance to hygromycin or GUS activity were obtained.
- Maize varieties A188, F1 (A188 x Black Mexican Sweet), F1 (A188 x B73Ht), F1 (B73Ht x A188) and F1 P3732 were selected as the sample materials.
- the varieties of A188, Black Mexican Sweet and B73Ht were obtained from National Institute of Agrobiological Resources, Ministry of Agriculture, Forestry & Fisheries, and P3732 was obtained from IWATA RAKUNOU KYODOKUMIAI.
- Mature seeds were immersed in 70% ethanol for 1 minute and in 1% sodium hypochlorite for 5 minutes. The seeds were then washed three times with sterilized water and were placed on LS solid medium (inorganic salts and vitamins of Linsmaier and Skoog; Linsmaier, E. and Skoog, F. 1965; Physiol. Plant 18: 100 - 127 , 100 mg/l of casamino acid, 700 mg/l of proline, 20 g/l of sucrose and 2.3 g/l of Gelrite). After culturing the seeds at 25°C in the dark for 4 days, tissues with a length of about 0.1 x 0.3 mm containing the apex dividing tissues were cut out and used as samples.
- LS solid medium inorganic salts and vitamins of Linsmaier and Skoog; Linsmaier, E. and Skoog, F. 1965; Physiol. Plant 18: 100 - 127 , 100 mg/l of casamino acid, 700 mg
- Immature embryos were placed on LSD1.5 solid medium (inorganic salts and vitamins of Linsmaier and Skoog, 100 mg/l of casamino acid, 700 mg/l of proline, 1.5 mg/l of 2,4-D, 20 g/l of sucrose and 2.3 g/l of Gelrite). After culturing the embryos for 3 weeks, the formed calli originated from scutella were collected and used in the subsequent experiments.
- LSD1.5 solid medium inorganic salts and vitamins of Linsmaier and Skoog, 100 mg/l of casamino acid, 700 mg/l of proline, 1.5 mg/l of 2,4-D, 20 g/l of sucrose and 2.3 g/l of Gelrite.
- Colonies of the Agrobacterium strains obtained by culturing the Agrobacterium strains on AB medium containing hygromycin (50 mg/l) and kanamycin (50 mg/l) for 3 - 10 days were collected using a platinum loop and the cells were suspended in the modified AA medium described in Example 1. The cell population was adjusted to 3 x 10 9 - 5 x 10 9 cells/ml and the resultants were used for inoculation.
- the sample tissues were immersed in the above-described suspensions of the Agrobacterium strains for 3 - 10 minutes.
- the tissues were then transferred to modified LS solid medium (inorganic salts of Linsmaier and Skoog, vitamins of Murashige and Skoog; Murashige, T. and Skoog, F. 1962; Physiol. Plant. 15:473-497 , 0.1 mg/l of kinetin, 1.0 mg/l of casamino acid and 2.3 g/l of Gelrite) and were cultured at 25°C under illumination for 2 - 3 days. Thereafter, the tissues were washed with sterilized water containing 250 mg/l of cefotaxime and then continued to be cultured on LS solid medium containing the same concentration of cefotaxime.
- the calli were immersed in the above-described Agrobacterium suspensions for about 5 minutes and the resulting calli were transferred to 2N6 solid medium containing acetosyringone described in Example 1 at 25°C in the dark for 3 days to carry out the culture with the Agrobacterium strains.
- the calli were washed with sterilized water containing 250 mg/l of cefotaxime and then continued to be cultured on LSD1.5 solid medium containing the same concentration of cefotaxime and 30 mg/l of hygromycin, thereby carrying out the selection of transformed calli.
- the shoot apex tissues and the calli immediately after the culture with the Agrobacterium strains and the shoot apex tissues and the calli which were continuously cultured after the culture with the Agrobacterium strains were examined for their GUS activities by the method described in Example 1.
- the invention is further defined by the following aspects:
Abstract
Description
- The present invention relates to a method for transforming monocotyledons.
- Conventional methods for transforming monocotyledons include electroporation method, polyethylene glycol method (PEG method), particle gun method and so on.
- The electroporation method is a method in which protoplasts and the desired DNA are mixed, and holes are formed in the cell membranes by electric pulse so as to introduce the DNA into the cells, thereby transforming the cells. This method currently has the highest reproducibility of the conventional methods and various genes have been introduced into monocotyledons, especially into rice plants by this method (Toriyama K. et al., 1988; Bio/Technol. 6:1072-1074, Shimamoto K. et al., 1989; Nature 338:274-276, Rhodes C.A. et al., 1989; Science 240:204-207). However, this method has the problems that 1) it can be applied only to the plant species for which the system for regenerating plants from protoplasts has been established, 2) since it takes several months to regenerate plants from the protoplasts, a long time is required to obtain transformants, and that 3) since the culture period is long, the frequency of emergence of mutants during the culture is high accordingly, so that the probability of obtaining normal transformants is decreased.
- The PEG method is a method in which the desired gene and protoplasts are mixed and the mixture is treated with PEG, thereby introducing the gene into the protoplasts. This method is different from the electroporation method in that PEG is used instead of the electric pulse. The efficiency of introducing the gene is thought to be somewhat lower than the electroporation method. Although there is a report that transformants were obtained by this method, this method is not widely used. Since protoplasts are used, this method has the same problems as in the electroporation method (Zhang W. et al., 1988; Theor. Appl. Genet. 76:835-840, Datta S.K. et al., 1990; Bio/Technol. 8:736-740).
- The particle gun method is a method in which the desired gene is attached to fine metal particles, and the metal particles are shot into cells or tissues at a high speed, thereby carrying out the transformation. Thus, according to this principle, transformation may be performed on any tissues. Therefore, this method is effective for transforming the plant species for which the systems for regenerating plants from protoplasts have not been established. The efficiency of transformation varies depending on the selection after the gene was shot. There is no data which compare the efficiency of this method with that of the electroporation method (Gordon-Kamm W.J. et al., 1990; Plant Cell 2:603-618, Fromm M.E. et al., 1990; Bio/Technol. 8:833-839, Christou P. et al., 1991; Bio/Technol. 9:957-962).
- Other methods include 1) culturing seeds or embryos with DNA (Topfer R. et al., 1989; Plant Cell 1:133-139, Ledoux L. et al., 1974 Nature 249:17-21); 2) treatment of pollen tube (Luo and Wu 1988; Plant Mol. Biol. Rep. 6:165-), 3) liposome method (Caboche M. 1990; Physiol. Plant. 79:173-176, Gad A.E. et al., 1990:177-183) and 4) microinjection method (Neuhaus G. et al., 1987; Theor. Appl. Genet. 75:30-36). However, these methods have problems in the efficiency of transformation, reproducibility or applicability, so that these methods are not commonly used.
- On the other hand, a method for introducing a gene using the Ti plasmid of bacteria belonging to genus Agrobacterium as a vector is widely used for transforming dicotyledons such as tobacco, petunia, rape and the like. However, it is said that the hosts of the bacteria belonging to genus Agrobacterium are restricted to dicotyledons and that monocotyledons are not parasitized by Agrobacterium (De Cleene M. 1976; Bot. Rev. 42:389-466).
- As for transformation of monocotyledons by Agrobacterium, although transformation of asparagus (Bytebier B. et al., 1987: Proc. Natl. Acad. Sci. USA, 84:5345-5349) and of Dioscorea bulbifera (Schafew et al., 1987; Nature 327:529-532) has been reported, it is said that this method cannot be applied to other monocotyledons, especially to the plants belonging to family Gramineae (Potrykus I. 1990; Bio/Technol. 8:535-543).
- Grimsley et al. (1987: Nature 325:177-179) reported that T-DNA of Agrobacterium in which DNA of maize streak virus was inserted was inoculated to the apical meristem of maize plants and infection of the plants by maize streak virus was confirmed. Since the infected symptoms are not observed when merely the DNA of maize streak virus is inoculated, they interpreted the above-mentioned result as a piece of evidence showing that Agrobacterium can introduce DNA into maize. However, since it is possible that a virus replicates even if it is not incorporated into the nucleus genome, the result does not show that the T-DNA was incorporated into the nucleus. They subsequently reported that the infection efficiency is the highest when the virus is inoculated to the apical meristem in the shoot apex of the maize (Grimsley et al., 1988: Bio/Technol. 6:185-189), and that virC gene in the plasmid of Agrobacterium is indispensable to the infection (Grimsley et al., Mol. Gen. Genet. 217:309-316).
- Gould J. et al. (1991; Plant Physiol. 95:426-434) inoculated super-virulent Agrobacterium ERA1 having a kanamycin-resistant gene and a GUS gene to shoot apices of maize after injuring the shoot apices with a needle, and selected the shoot apex based on the resistance to kanamycin. As a result, plants having resistance to kanamycin were obtained. They confirmed by Southern blotting analysis that some of the seeds of the subsequent generation of the selected plants had the introduced gene (chimera phenomenon).
- Mooney P.A. et al., (1991; Plant Cell, Tissue, Organ Culture 25:209-218) tried to introduce kanamycin-resistant gene into embryos of wheat using Agrobacterium. The embryos were treated with an enzyme to injure the cell walls, and then Agrobacterium was inoculated. Among the treated calli, although very small number of calli which were assumed to be transformants grew, plants could not be regenerated from these calli. The existence of the kanamycin-resistant gene was checked by Southern blotting analysis. As a result, in all of the resistant calli, change in structure of the introduced gene was observed.
- Raineri et al. (1990; Bio/Technol. 8:33-38) inoculated super-virulent Agrobacterium A281 (pTiBo542) to 8 varieties of rice after injuring the scutella of the rice plants. As a result, growth of tumor-like tissues was observed in two varieties, Nipponbare and Fujisaka 5. Further, an Agrobacterium containing a plasmid having a T-DNA from which a hormone-synthesizing gene was removed and instead, a kanamycin-resistant gene and GUS gene were inserted therein was inoculated to embryos of rice. As a result, growth of kanamycin-resistant calli was observed. Although the expression of GUS gene was observed in these resistant calli, transformed plants could not be obtained from the calli. They interpreted these results as that the T-DNA was introduced into rice cells.
- Thus, although the experimental results which suggest that introduction of genes into the plants belonging to family Gramineae such as rice, maize and wheat can be attained by using Agrobacterium have been reported, fully convincing results have not been obtained about the reproducibility, introduction efficiency and about the confirmation of the introduction of the gene (Potrykus I. 1990; Bio/Technol. 8:535-543).
- As mentioned above, introduction of genes into the plants belonging to family Gramineae is now mainly carried out by the electroporation method. However, with this method, since protoplasts are used, a long time and much labor are required to obtain regenerated plants. Further, there is a danger that mutants may emerge at a high frequency due to the long culturing period. Still further, this method cannot be applied to the plants such as maize for which the system for regenerating plants from protoplasts has not been established. In view of this, as mentioned above, as for maize, it has been tried to use the apical meristem. However, the operation for isolating the apical meristem requires much labor and it is not easy to prepare apical meristem in a large amount.
- Accordingly, an objective of the present invention is to provide a method for transforming monocotyledons, with which the time required for obtaining regenerated plants from the time of transformation is shorter than that in the conventional methods, which can be generally applied even to the plants for which the systems for regenerating plants from protoplasts have not yet been established, and with which the preparation of the materials to be used is easy.
- The present inventors intensively studied the influences of the monocotyledonous plant tissues treated with Agrobacterium, treatment conditions of Agrobacterium, constitution of the binary vector and the like on the introduction efficiency of the genes to discover that cultured tissues of monocotyledons can be transformed by using Agrobacterium with drastically high efficiency and reproducibility and that by employing this method, the above-mentioned object can be attained, thereby completing the present invention.
- That is, the present invention provides a method for transforming a monocotyledon comprising transforming a cultured tissue during dedifferentiation process or a dedifferentiated cultured tissue of said monocotyledon with a bacterium belonging to genus Agrobacterium containing a desired gene.
- By the present invention, it was first attained to introduce foreign genes to monocotyledons such as plants belonging to family Gramineae including rice, maize, wheat, barley and the like with good reproducibility. Although methods for transforming monocotyledons using Agrobacterium are known, they are not established methods as mentioned above. In contrast, according to the present invention, by inoculating Agrobacterium to cultured tissues which have not been employed in the conventional methods, genes can be very easily ' introduced. In the present invention, since a cultured tissue such as a callus which can be easily prepared is used, the sample materials can be obtained more easily than the conventional method employing the apical meristem. Further, since cultured cells are transformed, the time required for regenerating plants is shorter than in cases where protoplasts are transformed, so that the frequency of mutation is decreased. Further, by employing a super binary vector, it was first attained to introduce genes with high efficiency into varieties which are difficult to culture such as a variety of rice. Still further, as will be described in the examples below, by employing an appropriate selection method after inoculation, the chimera phenomenon in which the desired gene is introduced chimerally can be decreased.
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Fig. 1 shows the structure of pTOK162 which is an example of the plasmid contained in Agrobacterium bacteria that may be employed in the method of the present invention, and shows the method for constructing a plasmid pTOK232 used in the examples of the present invention. - The monocotyledons which may be transformed by the method according to the present invention are not restricted and the method according to the present invention may be applied to any monocotyledons such as rice, maize, barley, wheat, asparagus and the like.
- The cultured tissue used in the method of the present invention is a cultured tissue during dedifferentiation process or a dedifferentiated cultured tissue. The term "cultured tissue during the dedifferentiation process" herein means a tissue which is obtained by culturing an explant on a medium containing a plant growth regulator such as an auxin or a cytokinin, which tissue is before the stage that a callus or an adventitious embryo-like tissue is formed. The term "dedifferentiated tissue" herein means a callus or an adventitious embryo-like tissue obtained by culturing an explant in a medium containing a plant growth regulator such as an auxin or a cytokinin. The cultured tissue employed in the present invention may be originated from any part of the plant. For example, cultured tissues originated from scutellum, shoot apex, root, immature embryo, pollen and anther can be employed. As the cultured tissue employed in the present invention, it is preferred to employ a cultured tissue during callus-formation process which is cultured for not less than 7 days after an explant is placed on a dedifferentiation-inducing medium, a callus or an adventitious embryo-like tissue. In the interim, it is best preferred to employ a callus or an adventive embryo-like tissue as the cultured tissue. Dedifferentiation-inducing media are well-known in the art. For example, a medium containing major inorganic salts and vitamins of N6 medium (Chu C.C. 1987; Proc. Symp. Plant Tissue Culture, Science Press Peking, pp.43-50), 2 mg/l of 2,4-D, 1 g/l of casamino acid, 30 g/l of sucrose and 2 g/l of Gelrite, or a medium containing inorganic slats and vitamins of LS medium (Linsmaier, E., and Skoog, F. 1965; Physiol. Plant 18:100-127), 100 mg/l of casamino acid, 700 mg/l of proline, 1.5 mg/l of 2,4-D, 20 g/l of sucrose and 2.3 g/l of Gelrite. It should be noted that the cultured tissue employed in the method of the present invention is not necessarily a callus, but suspension cells may also be employed.
- As the Agrobacterium used for the transformation, Agrobacterium which are employed for the transformation of dicotyledons can be employed. Many of these Agrobacterium contain a vector having a DNA region originated from the virulence region (vir region) of Ti plasmid originated from Agrobacterium tumefaciens. The gene encoding a character which is desired to be given to the plant is inserted in this vector, or exists in a separate plasmid and inserted into the Ti plasmid in vivo by homologous recombination or the like. The present inventors previously developed a vector containing a DNA region originated from the virulence region (vir region) of Ti plasmid pTiBo542 (Jin S. et al., 1987: J. Bacteriol. 169:4417-4425) contained in a super-virulent Agrobacterium tumefaciens A281 exhibiting extremely high transformation efficiency (Hood E.E. et al., 1984; Bio/Technol. 2:702-709, Hood E.E. et al., 1986; J. Bacteriol. 168:1283-1290, Komari T. et al., 1986; J. Bacteriol. 166:88-94, Jin S. et al., 1987; J. Bacteriol. 169:4417-4425, Komari T. 1989; Plant Science 60:223-229 ATCC37394) (Japanese Laid-open Patent Application (Kokai) No.
4-222527 - An example of such a super binary vector is pTOK162 (Japanese Laid-open Patent Application (Kokai) No.
4-222527 EP-A-504,869 U.S. Serial No. 07/854,844 ). The structure is shown inFig. 1 . This plasmid comprises a plasmid called pTOK154 which can replicate in both Escherichia coli and in Agrobacterium tumefaciens (pTOK154 is a plasmid containing T region, which was constructed by the method described below from a known plasmid pGA472 derived from the Ti plasmid and a known plasmid having a wide host spectrum called pVCK101), into which a Kpn I fragment (containing virB, virG and virC genes) with a size of 15.2 kb originated from the virulence region of pTiBo542 is inserted, the Kpn I fragment having been cloned. In pTOK154, between two border sequences of the T region, a kanamycin-resistant gene is inserted as a gene to be introduced into a monocotyledon. This is an example wherein the gene desired to be introduced into the monocotyledon is arranged in a plasmid having the cloned DNA fragment originated from the virulence region of pTiBo542. InFig. 1 , each reference symbol represents the following meaning: - SP: spectinomycin-resistant gene
- HPT: hygromycin-resistant gene
- NPT: kanamycin-resistant gene
- TC: tetracycline-resistant gene
- IG: intron GUS gene
- BR: right border sequence of T-DNA
- BL: left border sequence of T-DNA
- virB, virC, virG: vir regions originated from super-virulent Agrobacterium tumefaciens A281
- ORI: replication origin of ColE1
- COS: COS site of λ phage
- K: restriction enzyme Kpn I site
- H: restriction enzyme Hind III site
- The gene which is desired to be incorporated into the monocotyledon may be inserted into a restriction site in the T region of the above-described plasmid, and the desired recombinant plasmid may be selected depending on an appropriate selection marker such as drug resistance and the like which the plasmid has. However, if the vector, like pTOK162 shown in
Fig. 1 , is large and has a number of restriction sites, it is not always easy to insert the desired DNA in the T region of the vector. In such a case, the desired DNA can be inserted in the T region of pTOK162 by utilizing the in vivo homologous recombination (Herrera-Esterella L. et al, 1983; EMBO J. 2:987-995, Horch R. H. et al. Science 1984; 223:496-498) in the cells of Agrobacterium tumefaciens. That is, pTOK162 is preliminarily introduced into Agrobacterium tumefaciens and the plasmid pBR322 (or a similar plasmid) containing the desired gene is further introduced into Agrobacterium tumefaciens. Since pTOK162 has a region homologous with a region of pBR322, the pBR322 derivative containing the desired gene is inserted into pTOK162 by the genetic recombination via the homologous regions. Unlike pTOK162, pBR322 cannot replicate by itself in Agrobacterium tumefaciens. Therefore, pBR322 can only be alive in Agrobacterium tumefaciens in the inserted form in pTOK162 (the recombined pTOK162 and pBR322 is hereinafter designated "pTOK162::pBR322 derivative"). By selecting the transformants based on the selection markers (such as drug resistance) specific to each of pTOK162 and pBR322 derivative, Agrobacterium tumefaciens transformants containing pTOK162::pBR322 derivative may be obtained. The present inventors made a study by introducing various plasmids into Agrobacterium tumefaciens containing pTOK162, to discover that as the selection marker of the pBR322 derivative, spectinomycin-resistant gene (SP) originated from transposon Tn7 (De Greve H.H. et al., 1981; Plasmid 6:235-248) is suited. Thus, in cases where the desired gene has already been cloned into pBR322, by inserting SP gene into the plasmid, the desired DNA can be inserted in the T region of pTOK162 by homologous recombination in vivo in Agrobacterium tumefaciens. Alternatively, a plasmid containing the DNA from pBR322 and SP gene is first provided, and the desired gene may be inserted into this plasmid. In this case, by utilizing the border sequences of the T region, it is possible to arrange the kanamycin-resistant gene and the desired gene in separate T regions in pTOK162. When plants are transformed using the resistance to kanamycin as a marker, there is a substantial probability that both T regions are introduced, the introduction of the desired gene can be sufficiently attained. Further, in this case, since both T regions may be inserted into different chromosomes, it may be possible to subsequently segregate the desired gene from the kanamycin-resistant gene. - The gene which is desired to be introduced to monocotyledons is not restricted at all and may be any gene which can give a desired character. Examples of the desired gene include herbicide-resistant genes, antibiotic-resistant genes, virus coat protein genes for giving resistance to the virus, genes related to starch formation in albumen and the like, although the desired genes are not restricted thereto.
- As the host bacterium belonging to genus Agrobacterium, Agrobacterium tumefaciens may preferably be employed, although not restricted.
- Introduction of a plasmid into the bacterium belonging to genus Agrobacterium such as Agrobacterium tumefaciens can be carried out by a conventional method such as triple cross method of bacteria (Ditta G. et al., 1980; Proc. Natl. Acad. Sci. USA 77:7347-7351).
- Since the Agrobacterium prepared as mentioned above has a highly virulent DNA originated from pTOK162, transformation of monocotyledons can be attained with a high efficiency.
- It should be noted that in the method of the present invention, although the gene which is desired to be introduced into the monocotyledon is arranged between border sequences of the T region as in the prior art, the desired gene may be arranged in the Ti plasmid or in another plasmid in the Agrobacterium.
- The transformation of the cultured tissue of a monocotyledon by the Agrobacterium may be carried out by merely contacting the cultured tissue with the Agrobacterium. For example, a cell suspension of the Agrobacterium having a population density of 106 -1011 cells/ml is prepared and the cultured tissue is immersed in this suspension for 3 - 10 minutes. The resulting culture tissue is then cultured on a solid medium for several days together with the Agrobacterium. Alternatively, transformation may be carried out by adding the Agrobacterium to the culture medium of the cultured tissue and continuously culturing the cultured tissue together with the Agrobacterium. Thus, in the method of the present invention, the cultured tissue may be subjected to the transformation without pretreatment such as treating the cultured tissue with an enzyme or injuring the cultured tissue.
- After subjecting the cultured tissues to the transformation, it is preferred to select the transformed cells or transformed tissues in the state during dedifferentiation process or in the dedifferentiated state. This can be attained by culturing the treated tissues on a medium containing a plant growth regulator such as an auxin or a cytokinin, which contains a selectable marker such as hygromycin and an antibiotic against the Agrobacterium, that is employed for the selection by the selection marker.
- The selected cells or the selected tissues may be subjected to regeneration culture by a known method. By this, plants acquired the desired character by the transformation can be regenerated.
- The present invention will now be described by way of examples thereof. It should be noted, however, that the present invention is not restricted to the examples.
- Varieties Asanohikari, Tsukinohikari and Koshihikari, which are varieties of japonica rice were selected as samples.
- Mature seeds of rice were sterilized by being immersed in 70% ethanol for 1 minute and then in 1% sodium hypochlorite solution for 30 minutes. The seeds were then placed on 2N6 solid medium (inorganic salts and vitamins of N6 (Chu C.C., 1978; Proc. Symp. Plant Tissue Culture, Science Press Peking, pp.43-50), 1g/l of casamino acid, 2 mg/l of 2,4-D, 30 g/l of sucrose, 2 g/l of Gelrite). Scutella were removed from the seeds on Day 4 from the beginning of the culture on 2N6 solid medium and used as "scutellum" samples. On the other hand, after culturing the mature seeds for about 3 weeks, the formed calli originated from scutella were transferred to 2N6 medium and cultured therein for 4 - 7 days. The resulting calli were used as "scutellum callus" samples.
- Mature seeds of rice were sterilized by the above-described method and were placed on 1/2 N6 solid medium (half strength of major inorganic salts and minor salts of N6, vitamins of N6, 1 g/l of casamino acid, 20 g/l of sucrose and 2 g/l of Gelrite). From seedlings on Day 3 after germination , tissues of 2 - 3 mm length containing apex dividing tissues were cut out and used as samples.
- From the seedlings obtained by the method descried in (iii), tip portions of 5 - 10 mm length of the seed roots were cut out and used as "root segment" samples. On the other hand, these radicles were cultured on 2N6 solid medium for about 2 weeks to obtain calli, and these calli were used as "root callus" samples.
- The calli originated from scutella were transferred to AA liquid medium (major inorganic salts of AA, amino acids of AA and vitamins of AA (Toriyama and Hinata 1985; Plant Science 41:179-183), MS minor salts (Murashige and Skoog 1962; Physiol. Plant. 15:473-497), 0.5 g/l of casamino acid, 1 mg/l of 2,4-D, 0.2 mg/l of kinetin, 0.1 mg/l of gibberellin and 20 g/l of sucrose) and the cells were cultured therein at 25°C in the dark under shaking of 120 rpm to obtain suspended cultured cells. The medium was replaced with fresh medium every week.
- Hygromycin resistant gene (HPT) and β-D-glucuronidase (GUS) gene were inserted in the T-DNA region of Ti plasmid to obtain the following plasmids:
- A plasmid in which the GUS gene containing the first intron of the catalase gene of caster bean and a hygromycin-resistant gene were ligated (Nakamura et al., 1991; Plant Biotechnology II (Extra Issue of GENDAI KAGAKU, pp.123-132), presented from Dr. Nakamura of Nagoya University).
- The Cla I fragment (2.5 kb) of the spectinomycin-resistant gene originated from Tn7 were treated with Klenow fragment to blunt the ends. The resulting fragment was inserted in Sma I site of pUC19 to obtain a plasmid pTOK107 (5.2 kb) having ampicillin-registant and spectinomycin-resistant genes. The obtained pTOK107 was treated with Eco RI and Hind III and the obtained 2.5 kb fragment containing the spectinomysin-resistant gene was ligated to a Eco RI - Hind III fragment (2.7 kb) of pGA482 to obtain pTOK170 (5.2 kb) containing the spectinomycin-resistant gene and has Hind III site and Hpa I site.
- A vector pIG221 in which the first intron of catalase of caster bean and GUS gene are ligated to 35S promoter (Ohta S. et al., 1990; Plant Cell Physiol. 31: 805-813, presented by Dr. Nakamura of Nagoya University) was digested with Eco RI and the resultant was treated with Klenow fragment to blunt the ends. To the resultant, a Hind III linker (pCAAGCTTG, code 4660P commercially available from TAKARA SHUZO). A fragment containing 35S promoter and intron GUS was cut out by digesting the resulting vector with Hind III, and the fragment was inserted into the Hind III site of a plasmid pGL2 (J. Paszkowski, obtained from Friedrich Mieocher Institute) containing a hygromycin-resistant gene ligated to 35S promoter, to obtain pGL2-IG (7.6 kb). The above-mentioned plasmid pGL2 was obtained by inserting a hygromycin-resistant gene (Gritz L. and Davis J. 1983; Gene 25: 179 - 188) into pDH51 (Pietrazak et al., 1986; Nucleic Acids Research 14: 5857 - 5868). The fragment obtained by treating pTOK170 with Hpa I was ligated to a Pvu II fragment (5.2 kb) of pGL2-IG to obtain pTOK229 (10.1 kb).
- Insertion of the desired genes (hygromycin-resistant gene and intron GUS gene) into a super binary vector pTOK162 obtained by inserting virB, virC and virG genes of strongly virulent Agrobacterium tumefaciens A281 into the binary vector was carried out by homologus recombination. That is, since both vectors contain a region originated from an E. coli plasmid pBR322, in the bacterial cells selected by resistances to spectinomycin and kanamycin, only the plasmid generated by recombination of the both plasmids are contained. The plasmid obtained by the fact that the hygromycin-resistant gene and the intron GUS gene were inserted into the super binary vector is called pTOK232 (see
Fig. 1 ). - Strains LBA4404 and EHA101 in which T-DNA regions were deleted were used as the host bacteria. Strain LBA4404 has a helper plasmid PAL4404 (having a complete vir region), and is available from American Type Culture Collection (ATCC 37349). Strain EHA101 has a helper plasmid having the vir region originated from a strongly virulent Agrobacterium tumefaciens A281, and is available from Hood E.E. et al., 1986.
- The various binary vectors described in (2) were introduced into these two strains of Agrobacterium tumefaciens, and the strains described in the following were used for introducing the genes. The plasmids were introduced into the Agrobacterium strains by triple cross (Ditta G. et al., 1980; Proc. Natl. Acad. Sci. USA 77: 7347-7351).
- LBA4404(pTOK232)
- LBA4404(pIG121Hm)
- EHA101(pIG121Hm)
- Colonies obtained by culturing the Agrobacterium strains on AB medium (Drlica K.A. and Kado C.I. 1974; Proc. Natl. Acad. Sci. USA 71:3677-3681) containing hygromycin (50 µg/ml) and kanamycin (50 µg/ml) for 3 - 10 days were collected with a platinum loop and suspended in modified AA medium (same as the composition of the above-described AA medium except that concentrations of sucrose and glucose were changed to 0.2 M and 0.2 M, respectively, and that 100 µM of acetosyringone was added, pH 5.2). The cell population was adjusted to 3 x 109 - 5 x 109 cells/ml and the suspensions were used for inoculation.
- The sample tissues were washed with sterilized water and immersed in the above-described suspensions of Agrobacterium strains, respectively, for 3 - 10 minutes. Thereafter, the shoot apex samples were placed on N6S3 solid medium (1/2 N6 major inorganic salts, N6 minor salts, N6 vitamins, Chu C.C., 1978, AA amino acids (Toriyama and Hinata 1985), 1 g/l of casamino acid, 0.2 mg/l of NAA, 1.0 mg/l of kinetin and 3 g/l of Gelrite) containing 100 µM of acetosyringone, 10 g/l of glucose and 20 g/l of sucrose. The other tissue samples such as scutellum callus samples were cultured on 2N6 solid medium containing acetosyringone, glucose and sucrose in the same concentrations as mentioned above. The both culture was carried out at 25°C in the dark for 2 - 5 days. The resulting inoculated tissues were then washed with sterilized water containing 250 mg/l of cefotaxime and then continued to be cultured on the respective solid media containing the same concentration of cefotaxime as mentioned above.
- Immediately after the above-mentioned culture with the Agrobacterium strains, the tissues were immersed in 0.1 M phosphate buffer (pH 6.8) containing 0.1% TRITON X-100 at 37°C for 1 hour. After washing off the Agrobacterium strains with phosphate buffer, phosphate buffer containing 0.1 mM of 5-bromo-4-chloro-3-indolyl-β-D-gluconic acid and 20% methanol was added to the tissues. After incubation at 37°C for 24 hours, the number of blue-colored tissues were counted under a microscope and the percentages thereof based on the number of samples are described. In the judgment of the GUS activities of the plants assumed to be transformants after the selection treatment, leaves were collected from the plants and GUS staining was performed in the same manner. If the entire leave or the cut face of the leave is uniformly colored in blue, the plant was judged to be a positive plant, and if the leave or the cut face of the leave is chimerically stained, the plant was judged to be a chimera plant.
- Shoot apices cultured with the Agrobacterium strains for 5 days were cultured on N6S3 medium containing 250 mg/l of cefotaxime for 2 weeks. The grown shoot apex tissues were transplanted to N6S3 medium containing 40 mg/l of hygromycin and selection of the transformants was carried out.
- Scutella cultured with the Agrobacterium strains for 3 days were cultured on 2N6 medium containing 250 mg/l of cefotaxime for 1 week. Selection of transformed cells were then carried out on 2N6 medium containing 50 mg/l of hygromycin.
- Tissues cultured with the Agrobacterium strains for 3 days were cultured on 2N6 medium containing 250 mg/l of cefotaxime for 1 week. Hygromycin-resistant cultured tissues were selected by culturing the cultured tissues on 2N6 medium containing 50 mg/l of hygromycin for 3 weeks (primary selection). The obtained resistant tissues were further cultured on N6-12 medium (N6 inorganic salts, N6 vitamins, 2 g/l of casamino acid, 0.2 mg/l of 2,4-D, 0.5 mg/l of 6BA, 5 mg/l of ABA, 30 g/l of sorbitol, 20 g/l of sucrose and 2 g/l of Gelrite) containing 50 mg/l of hygromycin for 2 - 3 weeks (secondary selection), and the calli grown on this medium were transferred to a plant regeneration medium N6S3 containing 0, 20 or 50 mg/l of hygromycin. In all of the media used after the culture with Agrobacterium strains, cefotaxime was added to 250 mg/l.
- The cells cultured with the Agrobacterium strains for 5 days were cultured in 2N6 medium containing 250 mg/l of cefotaxime for 1 week, and then the selection of the transformed cells was carried out on 2N6 medium containing 50 mg/l of hygromycin.
- Seeds of the progeny of the transformants were sown in aqueous 400-fold Homai hydrate (Kumiai Kagaku Inc.) solution containing 70 mg/l of hygromycin and incubated therein at 25°C for 10 days, thereby examining the resistance to hygromycin. Twenty seeds of each plant of the progeny of the transformants were sown and cultured for about 3 weeks. From the obtained seedlings, leaves were collected and examined for the expression of GUS gene.
- From the primary transformants of varieties Asanohikari and Tsukinohikari, DNAs were extracted by the method of Komari et al. (Komari et al., 1989; Theoretical and Applied Genetics 77: 547-552), and the DNAs were treated with a restriction enzyme Hind III. The resulting fragment was subjected to detection of the introduced genes by Southern blotting analysis using the HPT gene as a probe. The length of the Hind III fragment containing the HPT gene as a probe is about 5.5 kb and the length of the DNA region from the Hind III site in the T-DNA in this region to the L border sequence is about 5.4 kb (
Fig. 1 ). The Southern blotting analysis was carried out in accordance with Molecular Cloning (Sambrook et al., 1989; Cold Spring Harbor Laboratory Press). Two GUS positive plants, two GUS negative plants and two hygromycin-resistant plants were picked up from each of the two lines of the progeny of transformants of Tsukinohikari and were subjected to the Southern blotting analysis in the same manner as mentioned above. - In order to confirm that Agrobacterium can introduce genes into cells of monocotyledons, various tissues of the rice variety Tsukinohikari were treated with Agrobacterium tumefaciens EHA101 having a super-virulent vir region, into which the binary vector (supra) containing the hygromycin-resistant gene and the GUS gene were introduced, and then the GUS activities were examined. The sample tissues were shoot apices, radicles, scutella, radicle calli, scutellum calli and suspended cultured cells. In cases where the tissues were not treated with the Agrobacterium strain, no tissues exhibited GUS activity indicated by blue color. On the other hand, in cases where the tissues were treated with Agrobacterium tumefaciens EHA101 (pIG121Hm), in all of the tissues except for radicles, expression of GUS was confirmed. The ratio of the number of the tissues showing blue color to the number of treated tissues was the highest in scutellum calli (Table 1). Further, the size of the tissues expressing GUS was also largest in scutellum calli. The tissues exhibited the second highest rate of introduction next to the scutellum calli were shoot apices. Further, while the scutellum calli and the suspension cells which are dedifferentiated tissues of scutella exhibited high introduction rate, the introduction rate in scutella was apparently lower. This suggests that genes are more easily introduced into tissues having high cell-dividing activities.
Table 1 Differences in Efficiency of Introduction of GUS Gene Depending on Sample Material (Variety:Tsukinohikari) Sample Tissue Number of GUS+ Tissues /Number of Sample Tissues(%) Size of GUS-stained Portion Based on Treated Tissue Non-Treated Group Treated Group Shoot Apex 0/ 30 (0) 109/157 (69) + + + Root Segment 0/ 20 (0) 0/ 30 (0) Root Callus 0/ 30 (0) 24/115 (21) + Scutellum 0/ 50 (0) 8/ 89 (9) + Scutellum Callus 0/141 (0) 312/395 (79) + + + Suspension Cells 0/232 (0) 61/247 (25) + + + : 1% or less, + + : 1~10%, + + + : 10% or more - It has been confirmed that the binary vector pIG121Hm used in this experiment does not express GUS gene in Agrobacterium cells because the intron of caster oil plant is inserted in the promoter of the GUS gene (Nakamura et al., 1991). Thus, from the results of the experiments described above in which the expression of the GUS gene after the culturing with Agrobacterium is used as an index, it was confirmed that genes can be introduced into rice cells by Agrobacterium.
- Selection of transformed tissues and transformed cells was carried out according to the resistance to hygromycin, using shoot apices, scutella, scutellum calli and suspension cells after the culture with the Agrobacterium strain. As a result, growth of transformants exhibiting resistance to hygromycin were observed in scutellum calli and the suspension cells (Table 2). Further, all of the selected cells expressed the GUS gene. Although the shoot apex tissues exhibited high rate of introduction of GUS gene after the culture with the Agrobacterium strain, after the selection by hygromycin, all tissues died and no tissues resistant to hygromycin were obtained. Although shoot apex is a tissue containing the apical meristem, in order that cells expressing a hygromycin-resistance keep to grow after the treatment for introducing the gene, it is necessary that the gene be introduced into the limited apical meristem. The reason why no resistant tissues were obtained in spite of the fact that a number of genes were introduced into the shoot apices after the culture with the Agrobacterium strain is thought to be that the probability that the gene is introduced in the vicinity of the apical meristem is low. Further, it is easily assumed that even if a gene is introduced in the vicinity of the apical meristem so that transformed cells are obtained, the possibility that the obtained plants exhibit chimeric properties is high. From these, it is thought that the transformation method utilizing shoot apex, which was reported by Gould et al (1991) has more technical difficulties and less reproducibility than the method utilizing a dedifferentiated tissue such as callus.
- While transformed cells were obtained from the scutellum calli and the suspension cells which were originated from scutella of mature seeds, resistant cells were not obtained from the scutellum samples. Although the introduction of the genes was tried by using injured scutella in accordance with the teachings by Raineri et al. (1990), the efficiency of introducing genes was not promoted and no transformed cells were obtained. In contrast, in cases where scutellum calli were used as the samples, transformed cells were obtained with good reproducibility and high frequency, without a treatment such as injuring the samples. From these, it is thought that cultured tissues which were dedifferentiated or which are during the dedifferentiation process are suited as the tissues subjected to transformation by Agrobacterium.
Table 2 Differences in Rate of Energence of Transformed Tissues and Cells Depending on Sample Material (Variety:Tsukinohikari) Sample Tissue Number of Hygromycin-resistant Tissues /Number of Treated Tissues (%) Non-Treated Group Treated Group Shoot Apex 0/ 20 (0) 0/ 77 (0) Scutellum 0/ 30 (0) 0/128 (0) Scutellum Callus 0/ 50 (0) 169/743 (23) Suspension Cells 0/250 (0) 22/254 (9) - There are large differences among varieties about the conditions for establishing cultured cells and for regenerating plants from the cultured cells (Mikami and Kinoshita 1988; Plant Cell Tissue Organ Cult. 12:311 - 314). It is said that Koshihikari is difficult to culture among the Japonica rices. On the other hand, Tsukinohikari employed in the preceding section is relatively easy to culture. When using the transformation method utilizing Agrobacterium, it is practically inconvenient if such differences among varieties exist. In order to clarify this point, the differences in the efficiencies of gene introduction between Koshihikari and Tsukinohikari which have different easiness to culture were examined. The sample tissues employed were scutellum calli and the Agrobacterium tumefaciens strains employed were EHA101(pIG121Hm) and LBA4404(pIG121Hm).
- While GUS activity was observed in not less than 90% of calli of Tsukinohikari in each experiment, the GUS activity was observed in Koshihikari at lower rates (Table 3). Thus, in cases where EHA101(pIG121Hm) or LBA4404(pIG121Hm) is used, there is a difference in the introduction efficiency between the varieties.
Table 3 Differences in Rate of Introduction of GUS Gene Depending on Agrobacterium Strain and Rice Variety Number of GUS+ Tissues / Number of Treated Tissues (%) Strain Variety Experiment LBA4404 (pIG121Hm) EHA101 (pIG121Hm) LBA4404 (pTOK232) Tsukinohikari 1 67/70 (96) 78/87 (90) 64/66(97) Tsukinohikari 2 72/86 (84) 68/73 (93) 82/82(100) Koshihikari 1 46/135(34) 43/116(37) 124/131(95) Koshihikari 2 28/107(26) 81/143(57) 102/103(99) - EHA101(pIG121Hm) has a helper plasmid containing the vir region of super-virulent Agrobacterium tumefaciens A281. LBA4404(pIG121Hm) has an ordinary vir region. On the other hand, although the vir region of the helper plasmid in LBA4404(pTOK232) is ordinary, a gene which is a part of the vir region of the super-virulent Agrobacterium tumefaciens A281 is contained in the binary vector. This binary vector is originated from pTOK162 and made it possible to transform at a very high rate dicotyledonous species which are difficult to transform (Saito Y. et al., 1992; Theor. Appl. Genet. 83:679-683). Thus, there is a possibility that the transformation efficiency is largely influenced by the existence a super-virulent vir region or by the manner of existence thereof. Thus, using the above-described three Agrobacterium strains whose vir regions are different, the efficiencies of introducing GUS gene were compared. The samples used were scutellum calli of Koshihikari and Tsukinohikari.
- Even with LBA4404(pIG121Hm) which does not have a super-virulent vir region, tissues exhibiting GUS activities were obtained in both varieties. However, in Koshihikari, the rate was as low as about 30%. With EHA101(pIG121Hm) having the super-virulent vir region in the helper plasmid, the introduction efficiency in Koshihikari was somewhat higher. With LBA4404(pTOK232) having the super-virulent vir region in the binary vector, GUS activities were observed in not less than 95% tissues even with Koshihikari as with Tsukinohikari (Table 3). Further, as for the area of blue-colored regions in each tissue, the area was the largest with LBA4404(pTOK232), which indicates a high introduction efficiency.
- Using the above-mentioned 3 strains, the selection rates of hygromycin-resistant calli after culturing scutellum calli of Tsukinohikari and Koshihikari with the Agrobacterium strains were compared. As for the rate of emergence of the resistant calli, LBA4404(pTOK232) exhibited the highest rate. No differences about the rate of selection were observed between the varieties (Table 4). With the strains LBA4404(pIG121Hm) and EHA101(pIG121Hm), the rates of selection were low. Especially, with Koshihikari which is difficult to culture, the rate of emergence of hygromycin-resistant calli was as low as about 2%. Thus, it is thought that LBA4404(pTOK232) having a part of the super-virulent vir gene in the binary vector is the best as the Agrobacterium strain used for transforming rice.
Table 4 Differences in Transformation efficiency Depending on Agrobacterium Strain (Scutellum Callus) Number of Hygromycin-resistant Callus / Number of Treated Callus (%) Strain Variety Experiment LBA4404 (pIG121Hm) EHA101 (pIG121Hm) LBA4404 (pTOK232) Tsukinohikari 1 91/338 (27) 139/301 (46) 169/305 (55) Tsukinohikari 2 59/421 (14) 66/425 (16) 110/360 (31) Tsukinohikari 3 10/521 ( 2) 174/644 (27) Tsukinohikari 4 20/349 ( 6) 100/349 (29) Koshihikari 1 6/269 (2) 65/283 (23) - The thus obtained resistant calli were subjected to secondary selection, and plants were regenerated from the selected resistant calli. A group in which hygromycin was not added to the N6S3 medium for regeneration was provided. In this group, a number of plants which did not exhibit GUS activity or which exhibited GUS activity chimerally emerged. However, in cases where hygromycin was added to the regeneration medium, the number of these plants largely decreased and the number of plants each of which exhibits GUS activity in the whole plant was increased (Table 5, Table 6 and Table 7). In cases where the tissue was not treated with Agrobacterium, no plants which exhibited resistance to hygromycin or GUS activity were obtained. Therefore, the plants each of which exhibited GUS activity in the whole plant, which was regenerated from the hygromycin-resistant callus, are considered as transformants.
Table 5 Expression of GUS Gene in Plants Regenerated from Hygromycin-resistant Calli (Variety:Asanohikari) Resistant Callus Number of Regenerated Plants Expression of GUS Gene Stably Positive Chimera Negative 1 26 25 1 0 2 8 7 1 0 (Hygromycin was added to culture medium until regeneration of plants.) Table 6 Expression of GUS Gene in Plants Regenerated from Hygromycin-resistant Calli (Variety:Tsukinohikari) Sample Strain Number of Lines Sample Hygromycin -resistant Calli Calli Yielded Regenerated Plants GUS Positive Regenerated Plants LBA4404(pIG121Hm) 3 1 1 EHA101(pIG121Hm) 20 17 10 LBA4404(pTOK232) 20 15 12 (Hygromycin was added to culture medium until regeneration of plants.) Table 7 Expression of GUS Gene in Plants Regenerated from Hygromycin-resistant Calli (Variety:Asanohikari) Sample Strain Number of Lines Sample Hygromycin -resistant Calli Calli Yielded Regenerated Plants GUS Positive Regenerated Plants LBA4404(pIG121Hm) 19 5 3 EHA101(pIG121Hm) 11 4 1 LBA4404(pTOK232) 19 11 11 (Hygromycin was added to culture medium until regeneration of plants.) - When cultivated in a green house, the thus obtained transformants exhibited normal growth and morphology and no plants exhibited characteristics of tetraploid or malformation. As for the fertility of the seeds, although some plants exhibited partial infertility or complete infertility, most plants exhibited substantially normal fertility.
- Fragments obtained by Hind III digesting the whole DNAs in the primary transformants were subjected to detection of the introduced gene by Southern blotting analysis using the HPT gene as a probe. As a result, in all of the tested plants, the existence of the introduced gene in a number of 1 to several copies was confirmed (Table 8 and Table 9). While the size of the Hind III fragment containing the HPT gene in plasmid pTOK232 is 5.5 kb, in all of the tested transformants, a band having a size larger than about 6 kb was observed. This demonstrates that the T-DNA was incorporated into the plant chromosomes. The fact that the size of the detected DNA fragment differed from plant to plant indicates that the site in which the HPT gene was introduced was different from transformant to transformant. Therefore, it was confirmed that the fact that the introduced gene was detected was not due to the bacteria remaining in the plants.
- The resistance to hygromycin of the progeny of the transformants was examined. With the seeds of Control plants, substantially no germination was observed or the growth after germination was severely inhibited. In contrast, many of the seeds obtained from the transformants exhibited normal germination and growth (Tables 8 and 9). These hygromycin-resistant plants also expressed the GUS gene. In many lines, concerning the expression of hygromycin resistance and the GUS gene, genetic segregation substantially in accordance with one factor segregation was observed. As for transformant lines 1-2 and 3-2 of Asanohikari in Table 8, from the segregation ratio, existence of 2 or more factors of introduced genes is expected. The results of Southern blotting analysis were also compatible with the two factor segregation. In the transformant 2-1 in Table 8, existence of two copies of the introduced genes was confirmed. One of these bands represents a fragment shorter than 5 kb, so that it is assumed that the T-DNA of an incomplete form was incorporated. It is assumed that because of this, this transformant exhibited segregation like one factor segregation about the hygromycin resistance in the progeny.
- As shown in Table 9, many of the transformants of Tsukinohikari exhibited one factor segregation about the hygromycin resistance and GUS gene in the progeny. However, Southern blotting analysis of the primary transformant revealed that most of the transformants contained a plurality of copies of the gene although some of them contained a single copy of the gene. The progeny of the transformant 18a which contained one copy of the introduced gene and the progeny of the transformant 16c which contained two copies of the introduced gene were subjected to Southern blotting analysis. Two plants each of the GUS positive, GUS negative and hygromycin-resistant plants of each line were subjected to the analysis. As a result, in all of the plants except for the GUS negative plants, the same bands as detected in the plants of the primary transformants were detected. Therefore, it was shown that the introduced genes were inherited to the subsequent generations. As for the line 16c having two copies of the introduced gene, the fact that both the progeny of GUS positive plants and hygromycin-resistant plants had the same two copies of the introduced genes suggests that a plurality of genes were incorporated into the same chromosome or the same locus.
- These results indicate that the genes introduced into rice plants by Agrobacterium were incorporated in the nuclei of the plant cells and were inherited to the subsequent generations in accordance with the Mendel's laws.
Table 8 Number of Copies of Introduced Genes in Transformants Determined by Southern Blotting Analysis and Expression of Introduced Gene in the Progeny of Transformants (Variety:Asanohikari) Transformants Number of Copies of Introduced Gene Number of Plants of the Progeny of Transformants Resistance to Hygromycin Expression of GUS Resitant Sensitive Positive Negative Control - 0 60 0 20 1 - 2 2 30 0 19 1 2 - 1 2* 64 26 13 5 3-2 2 59 1 19 1 *In one of the two copies of the introduced genes, the restriction fragment was short, so that the introduced gene was incomplete. Table 9 Number of Copies of Introduced Genes in Transformants Determined by Southern Blotting Analysis and Expression of Introduced Gene in the Progeny of Transformants (Variety:Tsukinohikari) Transformants Number of Copies of Introduced Gene Number of Plants of the Progeny of Transformants Resistance to Hygromycin Expression of GUS Resitant Sensitive Positive Negative Control - 0 60 0 20 1 1 46 26 15 5 2a 2 33 18 13 5 2b 2 31 9 15 5 3 2 29 10 16 3 4a 3 22 21 13 7 4b 3 48 11 16 3 5a 3 26 13 17 3 5b 3 36 14 17 3 5c 3 24 9 17 2 6 2 47 13 - - 7 1 56 20 14 5 8 4 45 22 - - 9 1 52 18 18 2 10 4 53 10 - - 11 2 75 15 18 2 12 3 44 7 14 6 13a 2 33 18 15 5 13b 2 32 8 13 7 14a 1 72 20 15 5 14b 1 26 14 10 10 15 1~2 22 7 12 8 16a 2 31 10 15 2 16b 2 32 8 14 3 16c* 2 69 24 13 7 17 6 89 41 - - 18a* 1 35 5 15 5 18b 1 70 20 10 10 19 2 47 13 - - * Introduced genes in plants of the next generation were analyzed by Southern blotting analysis. - Maize varieties A188, F1 (A188 x Black Mexican Sweet), F1 (A188 x B73Ht), F1 (B73Ht x A188) and F1 P3732 were selected as the sample materials. The varieties of A188, Black Mexican Sweet and B73Ht were obtained from National Institute of Agrobiological Resources, Ministry of Agriculture, Forestry & Fisheries, and P3732 was obtained from IWATA RAKUNOU KYODOKUMIAI.
- Mature seeds were immersed in 70% ethanol for 1 minute and in 1% sodium hypochlorite for 5 minutes. The seeds were then washed three times with sterilized water and were placed on LS solid medium (inorganic salts and vitamins of Linsmaier and Skoog; Linsmaier, E. and Skoog, F. 1965; Physiol. Plant 18: 100 - 127, 100 mg/l of casamino acid, 700 mg/l of proline, 20 g/l of sucrose and 2.3 g/l of Gelrite). After culturing the seeds at 25°C in the dark for 4 days, tissues with a length of about 0.1 x 0.3 mm containing the apex dividing tissues were cut out and used as samples.
- Immature embryos were placed on LSD1.5 solid medium (inorganic salts and vitamins of Linsmaier and Skoog, 100 mg/l of casamino acid, 700 mg/l of proline, 1.5 mg/l of 2,4-D, 20 g/l of sucrose and 2.3 g/l of Gelrite). After culturing the embryos for 3 weeks, the formed calli originated from scutella were collected and used in the subsequent experiments.
- Among the strains of Agrobacterium described in Example 1, LBA4404(pTOK232) and EHA101(pIG121Hm) were used.
- Colonies of the Agrobacterium strains obtained by culturing the Agrobacterium strains on AB medium containing hygromycin (50 mg/l) and kanamycin (50 mg/l) for 3 - 10 days were collected using a platinum loop and the cells were suspended in the modified AA medium described in Example 1. The cell population was adjusted to 3 x 109 - 5 x 109 cells/ml and the resultants were used for inoculation.
- After piercing the cut out tissues with a glass needle, the sample tissues were immersed in the above-described suspensions of the Agrobacterium strains for 3 - 10 minutes. The tissues were then transferred to modified LS solid medium (inorganic salts of Linsmaier and Skoog, vitamins of Murashige and Skoog; Murashige, T. and Skoog, F. 1962; Physiol. Plant. 15:473-497, 0.1 mg/l of kinetin, 1.0 mg/l of casamino acid and 2.3 g/l of Gelrite) and were cultured at 25°C under illumination for 2 - 3 days. Thereafter, the tissues were washed with sterilized water containing 250 mg/l of cefotaxime and then continued to be cultured on LS solid medium containing the same concentration of cefotaxime.
- The calli were immersed in the above-described Agrobacterium suspensions for about 5 minutes and the resulting calli were transferred to 2N6 solid medium containing acetosyringone described in Example 1 at 25°C in the dark for 3 days to carry out the culture with the Agrobacterium strains. The calli were washed with sterilized water containing 250 mg/l of cefotaxime and then continued to be cultured on LSD1.5 solid medium containing the same concentration of cefotaxime and 30 mg/l of hygromycin, thereby carrying out the selection of transformed calli.
- The shoot apex tissues and the calli immediately after the culture with the Agrobacterium strains and the shoot apex tissues and the calli which were continuously cultured after the culture with the Agrobacterium strains were examined for their GUS activities by the method described in Example 1.
- In order to confirm that transformation employing the growth point tissues (shoot apex tissues) reported by Gould et al (Gould J. et al., 1991; Plant Physiol. 95:426-434) can be attained, isolated shoot apex tissues were treated with the above-described Agrobacterium tumefaciens strain EHA101(pIG121Hm), and the GUS activities of the grown plants were examined. While expression of the GUS gene was not observed in the tissues which were not treated with the Agrobacterium strain, expression of the GUS gene was observed in the spots pierced with the needle in the tissues which were treated with the Agrobacterium strain. The plants obtained by culturing the tissues were tested for GUS activities. However, no plants exhibited GUS activity. The vicinity of the growth point is a very small tissue, so that it is not easy to pierce the needle into the very small tissue to infect Agrobacterium. The results of this experiment show that the transformation by infecting the vicinity of the growth point with Agrobacterium requires high skill in cutting out and piercing the growth point and the like.
Table 10 Introduction of Gene into Maize Shoot Apex Tissue Experiment Number of Sample Tissues Number of Tissues whose shoot Apices Extended Number of Plants 0btained Number of Plants in which GUS was Expressed 1 24 9 2 0 2 16 8 6 0 3 17 13 5 0 4 14 1 0 0 5 45 14 7 0 6 32 14 8 0 7 30 7 1 0 Sample variety was P3732 in all experiments. - In any of the varieties tested, expression' of the GUS gene was observed at a high frequency. No differences were observed between the EHA101(pIG121Hm) and LBA4404(pTOK232) (Table 10). The size of the stained area with respect to the entire callus was not less than 10% in many calli, so that the expression of the gene was observed in wide range of cells. The binary vectors pIG121Hm and pTOK232 used in this experiment do not express GUS gene in Agrobacterium cells because the intron of caster bean is inserted in the promoter of the GUS gene. Therefore, the expression of the GUS gene observed in the maize calli indicates that gene introduction was carried out by the Agrobacterium strains with high frequency. After the culture with the Agrobacterium strains, from some of the sample calli, compact and knot-like calli grown. Since the grown cells exhibited expression of the GUS gene, it is thought that these cells are transformed cells. These compact and knot-like transformed calli can be regenerated to plants according to the method of Lupotto et al (Lupotto, E. and Lusardi, M.C. 1988; Maydica XXXIII:163-177).
Table 11 Efficiency of Introduction of GUS Gene into Maize Calli Variety Strain Number of GUS+ Calli / Number of Treated Calli (%) A188 1 32/35 (91) A188 1 34/34 (100) A188×BMS 1 41/49 (84) A188xB73 1 35/42 (83) A188 2 39/40 (98) A188 2 40/40 (100) A188×BMS 2 38/40 (95) A188xB73 2 31/40 (78) B73xA188 2 29/35 (83) BMS : Black Mexican Sweet
Strain 1 : EHA101(pIG121Hm), 2: LBA4404(pTOK232) - The invention is further defined by the following aspects:
- Aspect 1. A method for transforming a monocotyledon comprising transforming a cultured tissue during dedifferentiation process or a dedifferentiated cultured tissue of said monocotyledon with a bacterium belonging to genus Agrobacterium containing a desired gene.
- Aspect 2. The method according to aspect 1, wherein said monocotyledon is a plant belonging to family Gramineae.
- Aspect 3. The method according to aspect 1, wherein said monocotyledon is rice.
- Aspect 4. The method according to aspect 1, wherein said monocotyledon is corn.
- Aspect 5. The method according to any one of aspects 1 - 4, wherein said bacterium belonging to genus Agrobacterium contains Ti plasmid or Ri plasmid, which bacterium has a plasmid containing a DNA fragment originated from the virulence region of a Ti plasmid pTiBo542 of Agrobacterium tumefaciens.
- Aspect 6. The method according to aspect 5, wherein said plasmid containing said DNA fragment is pTOK162 or a derivative thereof.
- Aspect 7. The method according to any one of aspects 1 - 6, wherein said bacterium belonging to genus Agrobacterium is Agrobacterium tumefaciens.
- Aspect 8. The method according to any one of aspects 1 - 7, wherein cell population of said bacterium belonging to genus Agrobacterium used for transformation is 106 - 1011 cells/ml.
- Aspect 9. The method according to any one of aspects 1 - 8, wherein said cultured tissue is subjected to transformation without a pretreatment in which said cultured tissue is treated with an enzyme or in which said cultured tissue is injured.
- Aspect 10. The method according to any one of aspects 1 - 9, further comprising a step of selecting a transformed cell or a transformed tissue during dedifferentiation process or in dedifferentiated state, after subjecting said cultured tissue to transformation.
- Aspect 11. The method according to any one of aspects 1 - 9, wherein said cultured tissue is a cultured tissue during callus formation process which is attained by culturing a explant on a differentiation medium for not less than 7 days.
- Aspect 12. The method according to any one of aspects 1 - 11, wherein said cultured tissue is a cultured tissue originated from a somatic cell of said monocotyledon.
- Aspect 13. The method according to any one of aspects 1 - 12, wherein said cultured tissue has an ability to regenerate a normal plant.
Claims (16)
- A method for transforming a monocotyledon comprising transforming a cultured tissue during dedifferentiation process or a dedifferentiated cultured tissue of said monocotyledon with a bacterium belonging to genus Agrobacterium containing a desired gene,
wherein when the cultured tissue is rice, containing scutellem calli, the cultured tissue is not transformed with Agrobacterium tumefaciens strain LBA4404 (ATCC Accession No. 37349), the strain containing a pTOK232 plasmid containing the desired gene. - A method according to claim 1, wherein said monocotyledon is a plant belonging to family Gramineae.
- A method according to claim 1, wherein said monocotyledon is rice.
- A method according to claim 1, wherein said monocotyledon is corn.
- A method according to any preceding claim, wherein said bacterium belonging to genus Agrobacterium contains Ti plasmid or Ri plasmid, which bacterium has a plasmid containing a DNA fragment originated from the virulence region of a Ti plasmid pTiBo542 of Agrobacterium tumefaciens.
- A method according to claim 5, wherein said plasmid containing said DNA fragment is pTOK162 or a derivative thereof.
- A method according to any preceding claim, wherein said bacterium belonging to genus Agrobacterium is Agrobacterium tumefaciens.
- A method according to any preceding claim, wherein cell population of said bacterium belonging to genus Agrobacterium used for transformation is 106 - 1011 cells/ml.
- A method according to any preceding claim, wherein said cultured tissue is subjected to transformation without a pretreatment in which said cultured tissue is treated with an enzyme or in which said cultured tissue is injured.
- A method according to any preceding claim, further comprising a step of selecting a transformed cell or a transformed tissue during dedifferentiation process or in dedifferentiated state, after subjecting said cultured tissue to transformation.
- A method according to any one of claims 1 - 9, wherein said cultured tissue is a cultured tissue during callus formation process which is attained by culturing a explant on a differentiation medium for not less than 7 days.
- A method according to any preceding claim, wherein said cultured tissue is a cultured tissue originated from a somatic cell of said monocotyledon.
- A method according to any preceding claim, wherein said cultured tissue has an ability to regenerate a normal plant.
- A method according to any preceding claim, wherein the tissue is co-cultured on a solid medium together with Agrobacterium.
- A method according to any preceding claim, further comprising regenerating a plant having the introduced gene from the transformed tissue.
- A plant transformed by the method according to any preceding claim.
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US7705215B1 (en) * | 1990-04-17 | 2010-04-27 | Dekalb Genetics Corporation | Methods and compositions for the production of stably transformed, fertile monocot plants and cells thereof |
JP3209744B2 (en) | 1990-01-22 | 2001-09-17 | デカルブ・ジェネティクス・コーポレーション | Transgenic corn with fruiting ability |
US6777589B1 (en) | 1990-01-22 | 2004-08-17 | Dekalb Genetics Corporation | Methods and compositions for the production of stably transformed, fertile monocot plants and cells thereof |
US6329574B1 (en) * | 1990-01-22 | 2001-12-11 | Dekalb Genetics Corporation | High lysine fertile transgenic corn plants |
US6025545A (en) * | 1990-01-22 | 2000-02-15 | Dekalb Genetics Corporation | Methods and compositions for the production of stably transformed, fertile monocot plants and cells thereof |
US6395966B1 (en) * | 1990-08-09 | 2002-05-28 | Dekalb Genetics Corp. | Fertile transgenic maize plants containing a gene encoding the pat protein |
US6605712B1 (en) * | 1990-12-20 | 2003-08-12 | Arch Development Corporation | Gene transcription and ionizing radiation: methods and compositions |
US7060876B2 (en) * | 1992-07-07 | 2006-06-13 | Japan Tobacco Inc. | Method for transforming monocotyledons |
US6118047A (en) * | 1993-08-25 | 2000-09-12 | Dekalb Genetic Corporation | Anthranilate synthase gene and method of use thereof for conferring tryptophan overproduction |
US6326527B1 (en) * | 1993-08-25 | 2001-12-04 | Dekalb Genetics Corporation | Method for altering the nutritional content of plant seed |
US5780709A (en) * | 1993-08-25 | 1998-07-14 | Dekalb Genetics Corporation | Transgenic maize with increased mannitol content |
US6281411B1 (en) | 1993-08-25 | 2001-08-28 | Dekalb Genetics Corporation | Transgenic monocots plants with increased glycine-betaine content |
DK0672752T3 (en) | 1993-09-03 | 2004-10-04 | Japan Tobacco Inc | Method for transforming monocotyledons using scutella of undeveloped embryos |
CA2150475C (en) * | 1993-09-30 | 2009-02-24 | Jun Ueki | Phospholipase d gene originated from plant |
WO1995011979A1 (en) * | 1993-10-29 | 1995-05-04 | Japan Tobacco Inc. | Dna coding for carbonic anhydrase |
WO1995016031A1 (en) * | 1993-12-08 | 1995-06-15 | Japan Tobacco Inc. | Method of transforming plant and vector therefor |
JP3521161B2 (en) * | 1994-07-09 | 2004-04-19 | 日本たばこ産業株式会社 | DNA encoding phosphoenolpyruvate carboxykinase, recombinant vector containing the same, and transformed plant |
US6350934B1 (en) | 1994-09-02 | 2002-02-26 | Ribozyme Pharmaceuticals, Inc. | Nucleic acid encoding delta-9 desaturase |
US5631152A (en) * | 1994-10-26 | 1997-05-20 | Monsanto Company | Rapid and efficient regeneration of transgenic plants |
US6114608A (en) | 1997-03-14 | 2000-09-05 | Novartis Ag | Nucleic acid construct comprising bacillus thuringiensis cry1Ab gene |
JP3256952B2 (en) | 1994-11-09 | 2002-02-18 | 日本製紙株式会社 | Vector for gene transfer into plant, method for producing transgenic plant using the same, and method for multiple gene transfer into plant |
ID16389A (en) | 1996-03-28 | 1997-09-25 | Nippon Paint Co Ltd | METHOD OF CHANGING ORCHIDACEOUS PLANTS WITH AGROBACTERIUM |
EA199800212A1 (en) * | 1996-06-21 | 1998-10-29 | Монсанто Компани | METHODS OF OBTAINING SUSTAINABLE TRANSFORMABLE HIGH-PRODUCTIVE WHEAT BY TRANSFORMATION MEDIATED BY AGROBACTERIUM AND THE COMBINATION OBTAINED BY THEM |
US5773269A (en) * | 1996-07-26 | 1998-06-30 | Regents Of The University Of Minnesota | Fertile transgenic oat plants |
JPH10117776A (en) | 1996-10-22 | 1998-05-12 | Japan Tobacco Inc | Transformation of indica rice |
US5981840A (en) * | 1997-01-24 | 1999-11-09 | Pioneer Hi-Bred International, Inc. | Methods for agrobacterium-mediated transformation |
ID21006A (en) | 1997-02-10 | 1999-04-08 | Japan Tobacco Inc | C4 CYCLE TYPE PCK |
JP4199312B2 (en) * | 1997-02-20 | 2008-12-17 | バイエル・バイオサイエンス・エヌ・ブイ | Improved transformation of plants |
US6369298B1 (en) * | 1997-04-30 | 2002-04-09 | Pioneer Hi-Bred International, Inc. | Agrobacterium mediated transformation of sorghum |
US6162965A (en) * | 1997-06-02 | 2000-12-19 | Novartis Ag | Plant transformation methods |
EP2000538A3 (en) | 1997-06-03 | 2010-06-23 | The University of Chicago | Plant artificial chromosome (PLAC) compositions and methods for using them |
US7193128B2 (en) * | 1997-06-03 | 2007-03-20 | Chromatin, Inc. | Methods for generating or increasing revenues from crops |
US6183959B1 (en) | 1997-07-03 | 2001-02-06 | Ribozyme Pharmaceuticals, Inc. | Method for target site selection and discovery |
US6133035A (en) * | 1997-07-16 | 2000-10-17 | Dna Plant Technology Corporation | Method of genetically transforming banana plants |
US7087420B1 (en) | 1997-07-17 | 2006-08-08 | Cambia | Microbial β-glucuronidase genes, gene products and uses thereof |
US6391547B1 (en) * | 1997-09-09 | 2002-05-21 | Center For The Application Of Molecular Biology To International Agriculture | Microbial β-glucuronidase genes, gene products and uses thereof |
US6291244B1 (en) * | 1997-07-25 | 2001-09-18 | Sapporo Breweries Limited | Method of producing transformed cells of barley |
US6040498A (en) * | 1998-08-11 | 2000-03-21 | North Caroline State University | Genetically engineered duckweed |
CN1876819B (en) * | 1997-08-12 | 2010-06-23 | 北卡罗莱纳州立大学 | Genetically engineered duckweed |
US7161064B2 (en) * | 1997-08-12 | 2007-01-09 | North Carolina State University | Method for producing stably transformed duckweed using microprojectile bombardment |
US6586658B1 (en) | 1998-03-06 | 2003-07-01 | Metabolix, Inc. | Modification of fatty acid metabolism in plants |
US6437223B1 (en) | 1998-03-13 | 2002-08-20 | Syngenta Participations Ag | Inbred maize line 2070BT |
JP4474496B2 (en) | 1998-06-02 | 2010-06-02 | 学校法人日本大学 | IgA nephropathy-related DNA |
US6037522A (en) * | 1998-06-23 | 2000-03-14 | Rhone-Poulenc Agro | Agrobacterium-mediated transformation of monocots |
US7057090B1 (en) | 1998-07-17 | 2006-06-06 | Rutgers, The State University Of New Jersey | Agrobacterium-mediated transformation of turfgrass |
AR022383A1 (en) | 1998-09-18 | 2002-09-04 | Univ Kentucky Res Found | SYNTHESES |
WO2008021021A2 (en) | 2006-08-07 | 2008-02-21 | Mendel Biotechnology, Inc. | Plants with enhanced size and growth rate |
US20090265813A1 (en) | 2005-08-31 | 2009-10-22 | Mendel Biotechnology , Inc. | Stress tolerance in plants |
US7897843B2 (en) | 1999-03-23 | 2011-03-01 | Mendel Biotechnology, Inc. | Transcriptional regulation of plant biomass and abiotic stress tolerance |
US6420630B1 (en) | 1998-12-01 | 2002-07-16 | Stine Biotechnology | Methods for tissue culturing and transforming elite inbreds of Zea mays L. |
US6987025B1 (en) * | 1999-02-11 | 2006-01-17 | The Arizona Board Of Regents On Behalf Of The University Of Arizona | Dwf4 polynucleotides, polypeptides and uses thereof |
EP1586645A3 (en) * | 1999-02-25 | 2006-02-22 | Ceres Incorporated | Sequence-determined DNA fragments and corresponding polypeptides encoded thereby |
CA2267014A1 (en) * | 1999-03-26 | 2000-09-26 | Brenda Rojas | Monocot transformation using acrobacterium |
US7067719B1 (en) * | 1999-05-05 | 2006-06-27 | Seminis Vegetable Seeds, Inc. | Transformation of Allium sp. with Agrobacterium using embryogenic callus cultures |
US6429357B1 (en) | 1999-05-14 | 2002-08-06 | Dekalb Genetics Corp. | Rice actin 2 promoter and intron and methods for use thereof |
US7399850B2 (en) * | 1999-06-18 | 2008-07-15 | Ceres, Inc. | Sequence-determined DNA fragments encoding AP2 domain proteins |
JP4084502B2 (en) * | 1999-07-05 | 2008-04-30 | 独立行政法人科学技術振興機構 | Creation of iron deficiency resistant rice |
WO2001006844A1 (en) * | 1999-07-22 | 2001-02-01 | Japan As Represented By Director General Of National Institute Of Agrobiological Resources Ministry Of Agriculture, Forestry And Fisheries | Method for superrapid transformation of monocotyledon |
US6740526B1 (en) * | 1999-09-22 | 2004-05-25 | Wayne R. Curtis | Quantitative transient protein expression in plant tissue culture |
GB9923306D0 (en) | 1999-10-01 | 1999-12-08 | Isis Innovation | Diagnostic and therapeutic epitope, and transgenic plant |
WO2001033944A1 (en) | 1999-11-10 | 2001-05-17 | University Of Washington | Compositions and methods for modulation of plant cell division |
US20060194958A1 (en) * | 1999-11-10 | 2006-08-31 | Nickolai Alexandrov | Sequence-determined DNA fragments encoding AN1-like zinc finger proteins |
EP1780284B1 (en) | 1999-12-16 | 2009-10-28 | CropDesign N.V. | Optimized T-DNA transfer and vectors therefor |
JP4623825B2 (en) | 1999-12-16 | 2011-02-02 | 協和発酵バイオ株式会社 | Novel polynucleotide |
PT1255773E (en) | 1999-12-28 | 2009-04-28 | Bayer Bioscience Nv | Insecticidal proteins from bacillus thuringiensis |
CA2331674A1 (en) * | 2000-01-28 | 2001-07-28 | Southern Illinois University | Isolated polynucleotides and polypeptides relating to loci underlying resistance to soybean cyst nematode and soybean sudden death syndrome and methods employing same |
WO2001059091A2 (en) | 2000-02-11 | 2001-08-16 | Metabolix, Inc. | Multi-gene expression constructs containing modified inteins |
US6750379B2 (en) * | 2000-03-09 | 2004-06-15 | Dekalb Genetics Corporation | Homologous recombination-mediated transgene alterations in plants |
US6580019B1 (en) | 2000-03-09 | 2003-06-17 | Dekalb Genetics Corporation | Non-reciprocal recombination-mediated transgene deletion in transgenic plants |
US7691991B2 (en) * | 2000-04-17 | 2010-04-06 | Ceres, Inc. | Sequence-determined DNA fragments encoding cytochrome P450 proteins |
WO2001082685A1 (en) | 2000-04-28 | 2001-11-08 | Basf Aktiengesellschaft | Use of the maize x112 mutant ahas 2 gene and imidazolinone herbicides for selection of transgenic monocots, maize, rice and wheat plants resistant to the imidazolinone herbicides |
US7468475B2 (en) | 2000-06-16 | 2008-12-23 | Schmuelling Thomas | Method for modifying plant morphology, biochemistry and physiology |
EP1297163B1 (en) * | 2000-06-23 | 2011-04-27 | E.I. Du Pont De Nemours And Company | Recombinant constructs and their use in reducing gene expression |
US7902426B1 (en) * | 2000-08-03 | 2011-03-08 | Japan Tobacco Inc. | Method of improving gene transfer efficiency into plant cells utilizing heat and centrifugation |
CN1268752C (en) * | 2000-08-03 | 2006-08-09 | 日本烟草产业株式会社 | Method of improving gene transfer efficiency into plant cells |
JP2004506427A (en) | 2000-08-11 | 2004-03-04 | シンジェンタ パーティシペーションズ アクチェンゲゼルシャフト | Method for stable transformation of plants |
US7517975B2 (en) | 2000-09-26 | 2009-04-14 | Pioneer Hi-Bred International, Inc. | Nucleotide sequences mediating male fertility and method of using same |
CA2424602C (en) | 2000-10-06 | 2012-09-18 | Kyowa Hakko Kogyo Co., Ltd. | Antibody composition-producing cell |
FR2815969B1 (en) | 2000-10-30 | 2004-12-10 | Aventis Cropscience Sa | TOLERANT PLANTS WITH HERBICIDES BY METABOLIC BYPASS |
AU2002232554A1 (en) * | 2000-11-01 | 2002-05-27 | Maxygen, Inc. | Strawberry vein banding virus (svbv) promoter |
US6977293B1 (en) | 2000-11-03 | 2005-12-20 | Ceres, Inc. | Chimeric polypeptides |
DK2088200T3 (en) | 2000-11-17 | 2012-04-23 | Danisco Us Inc | Manipulation of the phenolic acid content and digestibility of plant cell walls by targeted expression of genes encoding cell wall degrading enzymes. |
ATE461273T1 (en) * | 2000-11-17 | 2010-04-15 | Metabolix Inc | PRODUCTION OF MEDIUM CHAIN POLYHYDROXYAL CANOATES FROM FATTY ACID BIOSYNTHESIS PATHWAYS |
AU2002214158B2 (en) | 2000-12-07 | 2007-01-18 | Syngenta Limited | Plant derived hydroxy phenyl pyruvate dioxygenases (HPPD) resistant against triketone herbicides and transgenic plants containing these dioxygenases |
US7385046B2 (en) * | 2001-01-03 | 2008-06-10 | Ceres, Inc. | Sequence-determined DNA fragments encoding ethylene responsive element binding proteins |
WO2002057407A2 (en) * | 2001-01-17 | 2002-07-25 | Avestha Gengraine Technologies Pvt. Ltd. | Novel method for transgenic plants by transformation & regeneration of indica rice plant shoot tips |
US8034791B2 (en) | 2001-04-06 | 2011-10-11 | The University Of Chicago | Activation of Egr-1 promoter by DNA damaging chemotherapeutics |
US20040242523A1 (en) * | 2003-03-06 | 2004-12-02 | Ana-Farber Cancer Institue And The Univiersity Of Chicago | Chemo-inducible cancer gene therapy |
WO2002080849A2 (en) * | 2001-04-06 | 2002-10-17 | University Of Chicago | Chemotherapeutic induction of egr-1 promoter activity |
AU2002256321A1 (en) | 2001-04-25 | 2002-11-05 | Cornell Research Foundation, Inc. | Tomato plants that exhibit resistance to $i(botrytis cinerea) |
WO2002092824A1 (en) * | 2001-05-15 | 2002-11-21 | Wilkinson Thesesa C | Method for producing transgenic monocotyledonous plants |
US7452705B2 (en) | 2001-05-22 | 2008-11-18 | The University Of Chicago | N4 virion single-stranded DNA dependent RNA polymerase |
US7193134B2 (en) | 2001-06-06 | 2007-03-20 | Bioriginal Food & Science Corp. | Flax (Linum usitatissimum L.) fatty acid desaturase |
EP1925671A1 (en) | 2001-06-06 | 2008-05-28 | Bioriginal Food & Science Corp. | Flax (Linum usitatissimum L.) seed-specific promoters |
EP1455568A2 (en) * | 2001-06-15 | 2004-09-15 | The University Of Toledo | Method for transformation of mono- and di- cotyledonous plants using meristematic tissue and nodal callus from dycotiledonous plants |
EP1279737A1 (en) | 2001-07-27 | 2003-01-29 | Coöperatieve Verkoop- en Productievereniging, van Aardappelmeel en Derivaten AVEBE B.A. | Transformation method for obtaining marker-free plants |
US7238862B2 (en) * | 2001-08-22 | 2007-07-03 | Monsanto Technology Llc | Efficiency Agrobacterium-mediated wheat transformation method |
CA2461844C (en) | 2001-09-27 | 2011-04-05 | Pioneer Hi-Bred International, Inc. | Phytate polynucleotides and methods of use |
ATE536100T1 (en) * | 2001-10-26 | 2011-12-15 | Baylor College Medicine | COMPOSITION FOR ALTERING BONE PROPERTIES IN A SUBJECT |
AU2002354015B2 (en) | 2001-11-07 | 2007-04-26 | Syngenta Participations Ag | Promoters for regulation of gene expression in plant roots |
KR100446987B1 (en) * | 2001-11-20 | 2004-09-01 | 학교법인 경희대학교 | Plant transformation vector pRCV2 containing hygromycin resistance gene for rescue cloning |
CN1615151A (en) * | 2001-12-11 | 2005-05-11 | 阿德维希斯公司 | Plasmid mediated suplementation for treating chronically ill subjects |
GB0130199D0 (en) | 2001-12-17 | 2002-02-06 | Syngenta Mogen Bv | New nematode feeding assay |
US20030166282A1 (en) * | 2002-02-01 | 2003-09-04 | David Brown | High potency siRNAS for reducing the expression of target genes |
US20060009409A1 (en) | 2002-02-01 | 2006-01-12 | Woolf Tod M | Double-stranded oligonucleotides |
EP1470148B1 (en) | 2002-02-01 | 2012-07-18 | Life Technologies Corporation | Double-stranded oligonucleotides |
US7135326B2 (en) * | 2002-02-07 | 2006-11-14 | E. I. Du Pont De Nemours And Company | UDP-glucosyltransferases |
US7534934B2 (en) | 2002-02-20 | 2009-05-19 | J.R. Simplot Company | Precise breeding |
NZ585768A (en) | 2002-02-20 | 2012-07-27 | Simplot Co J R | Precise plant breeding for the reduction of acrylamide |
US7598430B2 (en) * | 2002-03-20 | 2009-10-06 | J.R. Simplot Company | Refined plant transformation |
US20050034188A1 (en) * | 2002-03-20 | 2005-02-10 | J. R. Simplot Company | Refined plant transformation |
CA2911801A1 (en) | 2002-03-22 | 2003-10-02 | Greta Arnaut | Novel bacillus thuringiensis insecticidal proteins |
CA2481504C (en) | 2002-04-08 | 2011-08-23 | Pioneer Hi-Bred International, Inc. | Enhanced silk exsertion under stress |
US20030196219A1 (en) * | 2002-04-16 | 2003-10-16 | Monsanto Technology Llc | Novel Method for Agrobacterium Preparation for Plant Transformation |
EP1970442B1 (en) | 2002-05-03 | 2017-01-18 | Monsanto Technology LLC | Transgenic high tryptophan plants |
EP1549133B1 (en) | 2002-05-22 | 2015-04-22 | Monsanto Technology LLC | Fatty acid desaturases from fungi |
GB0212885D0 (en) | 2002-06-05 | 2002-07-17 | Isis Innovation | Therapeutic epitopes and uses thereof |
US20040248094A1 (en) * | 2002-06-12 | 2004-12-09 | Ford Lance P. | Methods and compositions relating to labeled RNA molecules that reduce gene expression |
US20040123350A1 (en) * | 2002-12-20 | 2004-06-24 | Ju-Kon Kim | A method for increasing an abiotic-resistance in monocot plant |
KR100440725B1 (en) * | 2002-06-20 | 2004-07-15 | 주식회사 그린진 바이오텍 | A Method for Increasing an Abiotic-Resistance in Monocot Plants |
US7329798B2 (en) * | 2002-06-28 | 2008-02-12 | University Of Guelph | Harvest-inducible regulatory elements and methods of using same |
AU2003245772A1 (en) | 2002-06-28 | 2004-01-19 | University Of Guelph | Harvest-inducible genes form alfalfa (medicago sativa) and methods of use thereof |
AU2003247650A1 (en) | 2002-06-28 | 2004-01-19 | Dow Agrosciences Llc | Pesticidally active proteins and polynucleotides obtainable from paenibacillus species |
US7364901B2 (en) | 2002-07-15 | 2008-04-29 | University Of Kentucky Research Foundation | Recombinant Stokesia epoxygenase gene |
US20060194959A1 (en) * | 2002-07-15 | 2006-08-31 | Nickolai Alexandrov | Sequence-determined DNA fragments encoding SRF-type transcription factors |
WO2005019409A2 (en) | 2002-07-15 | 2005-03-03 | Board Of Regents, The University Of Texas System | Combinatorial protein library screening by periplasmic expression |
AU2003304275A1 (en) | 2002-08-06 | 2005-01-21 | Pioneer Hi-Bred International, Inc. | Ap1 amine oxidase variants |
US20060064784A1 (en) | 2002-08-07 | 2006-03-23 | Basf Plant Science Gmbh | Nucleic acid sequences encoding proteins associated with abiotic stress response |
US20040029275A1 (en) * | 2002-08-10 | 2004-02-12 | David Brown | Methods and compositions for reducing target gene expression using cocktails of siRNAs or constructs expressing siRNAs |
US20060099224A1 (en) | 2002-08-12 | 2006-05-11 | David Kirn | Methods and compositions concerning poxviruses and cancer |
US7476777B2 (en) * | 2002-09-17 | 2009-01-13 | Ceres, Inc. | Biological containment system |
MXPA05003003A (en) * | 2002-09-17 | 2005-11-23 | Ceres Inc | Biological containment system. |
CA2501631A1 (en) | 2002-10-16 | 2004-04-29 | U.S. Smokeless Tobacco Company | Cloning of cytochrome p450 genes from nicotiana |
WO2004052085A1 (en) | 2002-12-06 | 2004-06-24 | Del Monte Fresh Produce Company | Transgenic pineapple plants with modified carotenoid levels and methods of their production |
US7078234B2 (en) | 2002-12-18 | 2006-07-18 | Monsanto Technology Llc | Maize embryo-specific promoter compositions and methods for use thereof |
JP4447467B2 (en) | 2002-12-26 | 2010-04-07 | 協和発酵バイオ株式会社 | Dipeptide production method |
EP1576178A4 (en) | 2002-12-26 | 2008-03-05 | Syngenta Participations Ag | Cell proliferation-related polypeptides and uses therefor |
BRPI0407514A (en) | 2003-02-17 | 2006-02-14 | Metanomics Gmbh | method for preparing a non-human organism, polynucleotide, method for preparing a vector, vector, host cell, polypeptide, antibody, antisense polynucleic acid, method for preparing a plant, plant cell, transgenic plant tissue, useful animal cell , useful animal or a transgenic microorganism non-human organism, transgenic microorganism, yield or a propagating material of a useful plant or animal, biomass of a microorganism, method for preparing fine chemicals, and, use of polypeptide |
CN1836045B (en) | 2003-03-28 | 2012-05-09 | 孟山都技术有限公司 | Novel plant promoters for use in early seed development |
US7612177B2 (en) | 2003-04-11 | 2009-11-03 | Cropdesign N.V. | Protein for use in modifying abiotic stress tolerance in yeast |
US20060288454A1 (en) | 2003-04-14 | 2006-12-21 | Sanz Molinero Ana I | Plants having modified growth characteristics and method for making the same |
ATE514781T1 (en) | 2003-04-15 | 2011-07-15 | Basf Plant Science Gmbh | NUCLEIC ACID SEQUENCES CODING FOR PROTEINS RELATED TO ABIOTIC STRESS RESPONSE, AS WELL AS PLANT CELLS AND PLANTS WITH INCREASED ENVIRONMENTAL STRESS TOLERANCE |
TW200424214A (en) * | 2003-04-21 | 2004-11-16 | Advisys Inc | Plasmid mediated GHRH supplementation for renal failures |
EP1620557A2 (en) | 2003-04-29 | 2006-02-01 | Pioneer Hi-Bred International, Inc. | Novel glyphosate-n-acetyltransferase (gat) genes |
EP1921152A1 (en) | 2003-05-05 | 2008-05-14 | Monsanto Technology, LLC | Transgenic plants with glycine-betaine specific promoter |
US7402734B2 (en) | 2003-06-16 | 2008-07-22 | Monsanto Technology Llc | Method and apparatus for preparation of genetically transformable plant tissue |
US7230161B2 (en) | 2003-06-23 | 2007-06-12 | Pioneer Hi-Bred International, Inc. | Engineering single-gene-controlled staygreen potential into plants utilizing ACC synthase from maize |
US7227414B2 (en) | 2003-06-27 | 2007-06-05 | Intel Corporation | Apparatus for receiver equalization |
AU2004262656A1 (en) | 2003-08-01 | 2005-02-17 | Basf Plant Science Gmbh | Process for the production of fine chemicals in plants |
US7150993B2 (en) * | 2003-08-05 | 2006-12-19 | Monsanto Technology Llc | Method for excision of plant embryos for transformation |
US7560611B2 (en) * | 2003-08-05 | 2009-07-14 | Monsanto Technology Llc | Method and apparatus for substantially isolating plant tissues |
CN101429522B (en) * | 2003-08-13 | 2011-08-31 | 日本烟草产业株式会社 | Method of elevating transformation efficiency in plant by adding copper ion |
WO2005017169A1 (en) * | 2003-08-13 | 2005-02-24 | Japan Tobacco Inc. | Method of infusing gene in plant material |
DK1656449T3 (en) | 2003-08-21 | 2009-06-02 | Monsanto Technology Llc | Fatty acid desaturases from primula |
PL2308961T3 (en) | 2003-08-25 | 2017-08-31 | Monsanto Technology Llc | Tubulin regulatory elements for use in plants |
AU2004270888B2 (en) | 2003-09-05 | 2008-08-07 | Cropdesign N.V. | Plants having modified growth characteristics and method for making the same |
AU2004278752B2 (en) | 2003-09-29 | 2008-04-24 | Monsanto Technology Llc | Methods for enhancing stress tolerance in plants and methods thereof |
WO2005033319A2 (en) | 2003-10-02 | 2005-04-14 | Monsanto Technology Llc | Stacking crop improvement traits in transgenic plants |
CN1867674B (en) | 2003-10-16 | 2011-11-09 | 美国无烟烟草有限责任公司 | Cloning of cytochrome P450 genes from nicotiana |
ES2346224T4 (en) | 2003-11-10 | 2011-06-01 | Icon Genetics Gmbh | PLANT EXPRESSION SYSTEM DERIVED FROM ARN VIRUSES. |
US20070169227A1 (en) | 2003-12-16 | 2007-07-19 | Pioneer Hi-Bred International Inc. | Dominant Gene Suppression Transgenes and Methods of Using Same |
EP1696721B1 (en) | 2003-12-16 | 2010-02-17 | Pioneer Hi-Bred International, Inc. | Dominant gene suppression transgenes and methods of using same |
EP1709188A1 (en) * | 2004-01-16 | 2006-10-11 | Ceres, Inc. | Plant cells having receptor polypeptides |
EP1718663B1 (en) | 2004-01-20 | 2011-07-13 | Monsanto Technology, LLC | Chimeric promoters for use in plants |
US7683237B2 (en) | 2004-02-10 | 2010-03-23 | Monsanto Technology Llc | Maize seed with synergistically enhanced lysine content |
AR047598A1 (en) | 2004-02-10 | 2006-01-25 | Monsanto Technology Llc | TRANSGENIZED CORN SEED WITH GREATER AMINO ACID CONTENT |
CN1950510B (en) | 2004-03-08 | 2011-10-05 | 先正达合作有限公司 | Glutamine-rich maize seed protein and promoter |
ATE517548T1 (en) | 2004-03-10 | 2011-08-15 | Monsanto Technology Llc | HERBICIDAL COMPOSITIONS CONTAINING N-PHOSPHONOMETHYLGLYCINE AND AN AUXIN HERBICIDE |
US20060041961A1 (en) | 2004-03-25 | 2006-02-23 | Abad Mark S | Genes and uses for pant improvement |
CN105002208A (en) | 2004-04-09 | 2015-10-28 | 孟山都技术有限公司 | Compositions and methods for control of insect infestations in plants |
ES2541537T3 (en) * | 2004-04-16 | 2015-07-21 | Monsanto Technology, Llc | Expression of fatty acid desaturases in corn |
AU2005235311B2 (en) | 2004-04-20 | 2009-10-08 | Syngenta Participations Ag | Regulatory sequences for expressing gene products in plant reproductive tissue |
CA2564807A1 (en) * | 2004-04-23 | 2005-11-10 | Ceres Inc. | Methods and materials for improving plant drought tolerance |
US10105437B2 (en) | 2004-04-28 | 2018-10-23 | Btg International Limited | Epitopes related to coeliac disease |
CN1976715B (en) | 2004-04-28 | 2014-07-30 | 英国技术集团国际有限公司 | Epitopes related to coeliac disease |
PT1740039E (en) | 2004-04-30 | 2012-08-27 | Dow Agrosciences Llc | Novel herbicide resistance genes |
CA2567764A1 (en) | 2004-06-16 | 2006-01-19 | Basf Plant Science Gmbh | Nucleic acid molecules encoding wrinkled1-like polypeptides and methods of use in plants |
US20050289667A1 (en) * | 2004-06-28 | 2005-12-29 | Cambia | Biological gene transfer system for eukaryotic cells |
WO2006069610A2 (en) | 2004-07-02 | 2006-07-06 | Metanomics Gmbh | Process for the production of fine chemicals |
EP1616959A1 (en) | 2004-07-07 | 2006-01-18 | Icon Genetics AG | Biological safe transient protein expression in plants |
BRPI0514003A (en) | 2004-07-31 | 2008-05-20 | Metanomics Gmbh | methods for preparing a non-human organism, preparing a vector, a transgenic plant, a plant cell, a plant tissue, a useful animal's cell, a useful animal or a transgenic microorganism, and fine chemicals, and for identifying a gene product, polynucleotide , vector, host cell, antibody, antisense nucleic acid, non-human organism, transgenic microorganism, seed, tuber or propagation material of a plant, biomass of a microorganism, plant cell, plant cell organelle, plant tissue, plant or part thereof , and, uses of a polypeptide, and nucleic acid molecule |
US20060075522A1 (en) | 2004-07-31 | 2006-04-06 | Jaclyn Cleveland | Genes and uses for plant improvement |
AU2005268917B2 (en) | 2004-08-02 | 2010-05-20 | Basf Plant Science Gmbh | Method for isolation of transcription termination sequences |
WO2006023766A2 (en) * | 2004-08-20 | 2006-03-02 | Ceres Inc. | P450 polynucleotides, polypeptides, and uses thereof |
WO2006023869A2 (en) | 2004-08-24 | 2006-03-02 | Monsanto Technology Llc | Adenylate translocator protein gene non-coding regulatory elements for use in plants |
US7595433B2 (en) | 2004-09-14 | 2009-09-29 | Ceres, Inc. | Modulations of amino acid and sugar content in plants |
BRPI0515302A (en) | 2004-09-14 | 2008-07-15 | Monsanto Technology Llc | promoter molecules for use in plants |
US20060059585A1 (en) * | 2004-09-14 | 2006-03-16 | Boris Jankowski | Modulating plant sugar levels |
CA2579927A1 (en) | 2004-09-24 | 2006-03-30 | Basf Plant Science Gmbh | Plant cells and plants with increased tolerance to environmental stress |
BRPI0515902A (en) | 2004-09-24 | 2008-08-12 | Basf Plant Science Gmbh | plant cell, plant, seed, isolated nucleic acid molecule, nucleic acid construct, vector, host cell, isolated strain-related protein, methods of producing a transgenic plant and modifying a plant's voltage tolerance, and, uses of a srp encoding nucleic acid and of increased tolerance and / or resistance to environmental stress and / or nucleic acid |
CA2582081C (en) * | 2004-10-01 | 2022-05-17 | De Ruiter Seeds R&D B.V. | Pmmov resistant capsicum plants |
US7429692B2 (en) * | 2004-10-14 | 2008-09-30 | Ceres, Inc. | Sucrose synthase 3 promoter from rice and uses thereof |
EP1807522B1 (en) | 2004-10-21 | 2013-11-20 | Venganza Inc. | Methods and materials for conferring resistance to pests and pathogens of plants |
EP1652930A1 (en) | 2004-10-25 | 2006-05-03 | De Ruiter Seeds R&D B.V. | Botrytis-resistant tomato plants |
WO2006137941A2 (en) | 2004-11-12 | 2006-12-28 | Ambion, Inc. | Methods and compositions involving mirna and mirna inhibitor molecules |
WO2007050625A1 (en) * | 2005-10-25 | 2007-05-03 | Ceres, Inc. | Modulation of triterpenoid content in plants |
EP1974049A2 (en) | 2004-12-17 | 2008-10-01 | Metanomics GmbH | Process for the control of production of fine chemicals |
AU2005323166B2 (en) | 2004-12-21 | 2011-11-10 | Monsanto Technology, Llc | Recombinant DNA constructs and methods for controlling gene expression |
US20060200878A1 (en) | 2004-12-21 | 2006-09-07 | Linda Lutfiyya | Recombinant DNA constructs and methods for controlling gene expression |
US20070199095A1 (en) | 2005-10-13 | 2007-08-23 | Edwards Allen | Methods for producing hybrid seed |
CN101442902B (en) | 2004-12-21 | 2013-09-18 | 孟山都技术有限公司 | Preparation method of transgenic plants with enhanced agronomic traits |
US8314290B2 (en) | 2004-12-21 | 2012-11-20 | Monsanto Technology Llc | Temporal regulation of gene expression by MicroRNAs |
EP3078749B1 (en) | 2004-12-21 | 2019-10-09 | Monsanto Technology LLC | Transgenic plants with enhanced agronomic traits |
AU2006204997B2 (en) | 2005-01-12 | 2011-09-01 | Monsanto Technology, Llc | Genes and uses for plant improvement |
EP2365086A1 (en) | 2005-02-09 | 2011-09-14 | Bioriginal Food & Science Corporation | Novel omega-3 fatty acid desaturase family members and uses thereof |
US20140199313A1 (en) | 2005-03-02 | 2014-07-17 | Metanomics Gmbh | Process for the Production of Fine Chemicals |
CA2598792A1 (en) | 2005-03-02 | 2006-09-08 | Metanomics Gmbh | Process for the production of fine chemicals |
EP2169058B1 (en) | 2005-03-08 | 2013-01-23 | BASF Plant Science GmbH | Expression enhancing intron sequences |
BRPI0608637A2 (en) | 2005-03-16 | 2010-01-19 | Metabolix Inc | recombinant vector for enzyme expression, transformed plant cell, method for biosynthetic product production |
EP2444496A1 (en) | 2005-04-15 | 2012-04-25 | Del Monte Fresh Produce Company | Plant promoters, terminators, genes, vectors and related transformed plants |
CA2604807C (en) | 2005-04-19 | 2018-06-12 | Basf Plant Science Gmbh | Improved methods controlling gene expression |
ATE552343T1 (en) | 2005-04-19 | 2012-04-15 | Basf Plant Science Gmbh | ENDOSPERM-SPECIFIC EXPRESSION AND/OR EXPRESSION IN GERMINATING EMBRYOS OF MONOCOTYLEDONE PLANTS |
WO2006122023A1 (en) | 2005-05-09 | 2006-11-16 | The Samuel Roberts Noble Foundation, Inc. | Agrobacterium transformation of stolons |
EP2478760A1 (en) | 2005-05-10 | 2012-07-25 | Monsanto Technology LLC | Genes and uses for plant improvement |
CN100360677C (en) * | 2005-05-13 | 2008-01-09 | 吉林省农业科学院 | Transgene method for plant |
NZ564016A (en) | 2005-05-25 | 2009-08-28 | Pioneer Hi Bred Int | Methods for improving crop plant architecture and yield by introducing recombinant expression cassette |
US8124839B2 (en) * | 2005-06-08 | 2012-02-28 | Ceres, Inc. | Identification of terpenoid-biosynthesis related regulatory protein-regulatory region associations |
US7994399B2 (en) | 2005-06-23 | 2011-08-09 | Basf Plant Science Gmbh | Methods for the production of stably transformed, fertile Zea mays plants |
ES2435079T3 (en) | 2005-07-08 | 2013-12-18 | Universidad Nacional Autonoma De Mexico Instituto | New bacterial proteins with pesticide activity |
BRPI0614695A2 (en) | 2005-07-29 | 2011-04-12 | Targeted Growth Inc | dominant negative mutant krp protein protection from inhibition of active cyclin-cdk complex by wild type krp |
NZ542110A (en) * | 2005-08-30 | 2008-07-31 | Horticulture & Food Res Inst | Compositions and methods for modulating pigment production in plants |
US8993846B2 (en) | 2005-09-06 | 2015-03-31 | Monsanto Technology Llc | Vectors and methods for improved plant transformation efficiency |
US8980246B2 (en) | 2005-09-07 | 2015-03-17 | Sillajen Biotherapeutics, Inc. | Oncolytic vaccinia virus cancer therapy |
JP2009507853A (en) * | 2005-09-07 | 2009-02-26 | ジェンネレックス インコーポレイティッド | Systemic treatment of metastatic and / or systemic disseminated cancer using GM-CSF expressing poxvirus |
CA2621874C (en) | 2005-09-08 | 2014-12-16 | Chromatin Inc. | Plants modified with mini-chromosomes |
US9121028B2 (en) * | 2005-09-09 | 2015-09-01 | Monsanto Technology Llc | Selective gene expression in plants |
ATE548459T1 (en) | 2005-09-16 | 2012-03-15 | Monsanto Technology Llc | METHOD FOR THE GENETIC CONTROL OF INSECT INFESTATION IN PLANTS AND COMPOSITIONS |
EP1931789B1 (en) | 2005-09-20 | 2016-05-04 | BASF Plant Science GmbH | Methods for controlling gene expression using ta-siran |
US8859846B2 (en) | 2005-09-21 | 2014-10-14 | E. I. Du Pont De Nemours And Company | Doubling of chromosomes in haploid embryos |
US20090270314A1 (en) | 2005-09-29 | 2009-10-29 | Tohoku University | Polypeptide having anti-angiogenic activity |
WO2007041536A2 (en) * | 2005-09-30 | 2007-04-12 | Ceres, Inc. | Modulating plant tocopherol levels |
ATE542912T1 (en) | 2005-10-03 | 2012-02-15 | Monsanto Technology Llc | TRANSGENIC PLANT SEEDS WITH INCREASED LYSINE |
EP1947925B1 (en) | 2005-10-20 | 2013-12-25 | Commonwealth Scientific And Industrial Research Organisation | Cereals with altered dormancy |
NZ567807A (en) | 2005-10-28 | 2011-09-30 | Dow Agrosciences Llc | Novel herbicide (2,4-D and pyridyloxyacetate) resistance genes with aryloxyalkanoage dioxygenase activity |
EP1782685A1 (en) * | 2005-11-04 | 2007-05-09 | De Ruiter Seeds R&D B.V. | Disease resistant cucumber plants |
CA2632405A1 (en) | 2005-12-09 | 2007-06-14 | Basf Plant Science Gmbh | Nucleic acid molecules encoding polypeptides involved in regulation of sugar and lipid metabolism and methods of use viii |
WO2007069301A1 (en) * | 2005-12-13 | 2007-06-21 | Japan Tobacco Inc. | Method of elevating transforming efficiency by using powder |
BRPI0619837A2 (en) | 2005-12-15 | 2011-10-18 | Targeted Growth Inc | method for increasing the seed size of a plant and / or for increasing the number of seeds obtainable from a plant, genetic construction and methods for producing a transgenic plant |
AU2005339717A1 (en) | 2005-12-21 | 2007-07-12 | Monsanto Technology, Llc | Transgenic plants with enhanced agronomic traits |
EP1800535A1 (en) | 2005-12-21 | 2007-06-27 | De Ruiter Seeds R&D B.V. | Closterovirus-resistant melon plants |
WO2008019162A2 (en) | 2006-01-18 | 2008-02-14 | University Of Chicago | Compositions and methods related to staphylococcal bacterium proteins |
CA2821436A1 (en) | 2006-02-09 | 2007-08-16 | Pioneer Hi-Bred International, Inc. | Genes for enhancing nitrogen utilization efficiency in crop plants |
BRPI0707626A2 (en) | 2006-02-10 | 2011-05-10 | Monsanto Technology Llc | identification and use of target genes for the control of plant parasitic nematodes |
CN101421406B (en) | 2006-02-13 | 2016-08-31 | 孟山都技术有限公司 | For producing nucleic acid construct and the method that the seed oil of change forms |
CN101384721A (en) | 2006-02-13 | 2009-03-11 | 孟山都技术有限公司 | Selecting and stabilizing dsrna constructs |
EP2843053A1 (en) | 2006-02-17 | 2015-03-04 | Monsanto Technology LLC | Chimeric regulatory sequences comprising introns for plant gene expression |
WO2007120989A2 (en) * | 2006-02-24 | 2007-10-25 | Ceres, Inc. | Shade regulatory regions |
BRPI0712914A2 (en) * | 2006-02-27 | 2012-10-09 | Edenspace System Corp | energy crops for improved biofuel feedstock |
EP2343375A3 (en) | 2006-03-24 | 2012-02-08 | BASF Plant Science GmbH | Proteins associated with abiotic stress response and homologs |
WO2008034648A1 (en) | 2006-04-05 | 2008-03-27 | Metanomics Gmbh | Process for the production of a fine chemical |
WO2007117693A2 (en) * | 2006-04-07 | 2007-10-18 | Ceres, Inc. | Regulatory protein-regulatory region associations related to alkaloid biosynthesis |
WO2007119796A1 (en) | 2006-04-14 | 2007-10-25 | Medical And Biological Laboratories Co., Ltd. | Mutant polypeptide having effector function |
AR060563A1 (en) | 2006-04-24 | 2008-06-25 | Mendel Biotechnology Inc | DISEASE PROMOTERS FOR DISEASE |
EP1849871A1 (en) | 2006-04-25 | 2007-10-31 | De Ruiter Seeds R&D B.V. | Tomato plants having higher levels of resistance to Botrytis |
MX295027B (en) | 2006-05-12 | 2012-01-20 | Monsanto Technology Llc | Methods and compositions for obtaining marker-free transgenic plants. |
JP2009537150A (en) | 2006-05-16 | 2009-10-29 | モンサント テクノロジー エルエルシー | Use of non-Agrobacterium species for plant transformation |
EP2455489A3 (en) | 2006-05-25 | 2012-07-11 | Monsanto Technology LLC | A method to identify disease resistant quantitative trait loci in soybean and compositions thereof |
WO2007137973A2 (en) | 2006-05-31 | 2007-12-06 | Metanomics Gmbh | Manipulation of the nitrogen metabolism using ammonium transporter or glucose 6-phosphate deshydrogenases or farnesyl phosphate synthetase (fpp) |
US7855326B2 (en) | 2006-06-06 | 2010-12-21 | Monsanto Technology Llc | Methods for weed control using plants having dicamba-degrading enzymatic activity |
CA2918565C (en) | 2006-06-29 | 2018-04-10 | Mendel Biotechnology, Inc. | Improved yield and stress tolerance in transgenic plants with overexpressing polypeptides comprising a b-box zinc-finger domain |
US7666677B2 (en) | 2006-07-05 | 2010-02-23 | Luis Fabricio Medina-Bolivar | Production of stilbenes in plant hairy root cultures |
US20080043087A1 (en) | 2006-07-19 | 2008-02-21 | James Cowan | Digital transfer method for printing on a target surface |
BR122017006335B1 (en) | 2006-07-19 | 2018-06-26 | Monsanto Technology Llc | METHOD OF INCREASING EFFICIENCY OF BACTERIA-MEDIATED PLANT TRANSFORMATION RECOMBINANT DNA CONSTRUCTION AND TRANSGENIC BACTERIAL CELL |
WO2008021207A2 (en) | 2006-08-11 | 2008-02-21 | Dow Agrosciences Llc | Zinc finger nuclease-mediated homologous recombination |
US20100293673A1 (en) | 2006-08-15 | 2010-11-18 | Jason Bull | Compositions and Methods of Plant Breeding Using High Density Marker Information |
EP2048939A4 (en) | 2006-08-17 | 2010-04-28 | Monsanto Technology Llc | Transgenic plants with enhanced agronomic traits |
CL2007002532A1 (en) * | 2006-08-31 | 2008-01-11 | Monsanto Technology Llc Soc Organizada Bajo Las Leyes Del Estado De Delaware | Procedure to identify genes that confer improved traits of a plant population. |
WO2008028121A1 (en) * | 2006-08-31 | 2008-03-06 | Monsanto Technology Llc | Plant transformation without selection |
US8404928B2 (en) | 2006-08-31 | 2013-03-26 | Monsanto Technology Llc | Phased small RNAs |
ES2551892T3 (en) | 2006-09-15 | 2015-11-24 | Ottawa Health Research Institute | Oncolytic rhabdovirus |
AU2007302306A1 (en) | 2006-09-25 | 2008-04-03 | Freie Universitat Berlin | Transcriptional repressors of cytokinin signaling and their use |
JP5256039B2 (en) | 2006-09-25 | 2013-08-07 | 協和発酵バイオ株式会社 | Dipeptide production method |
CN101802215A (en) | 2006-10-12 | 2010-08-11 | 孟山都技术有限公司 | Plant micrornas and using method thereof |
EP2121936A2 (en) | 2006-10-13 | 2009-11-25 | BASF Plant Science GmbH | Plants with increased yield |
AP2885A (en) | 2006-10-16 | 2014-05-31 | Monsanto Technology Llc | Methods and compositions for improving plant health |
US7939721B2 (en) | 2006-10-25 | 2011-05-10 | Monsanto Technology Llc | Cropping systems for managing weeds |
US8222388B2 (en) * | 2006-11-22 | 2012-07-17 | Ceres, Inc. | Broadly expressing regulatory regions |
US8067179B2 (en) | 2006-11-30 | 2011-11-29 | Research Development Foundation | Immunoglobulin libraries |
KR101613624B1 (en) | 2006-12-14 | 2016-04-29 | 다우 아그로사이언시즈 엘엘씨 | Optimized non-canonical zinc finger proteins |
US20110236374A1 (en) | 2007-01-24 | 2011-09-29 | Kyowa Hakko Kirin Co., Ltd. | Genetically recombinant antibody composition capable of binding specifically to ganglioside gm2 |
US7994290B2 (en) | 2007-01-24 | 2011-08-09 | Kyowa Hakko Kirin Co., Ltd | Effector function enhanced recombinant antibody composition |
JPWO2008090678A1 (en) | 2007-01-25 | 2010-05-13 | 協和発酵キリン株式会社 | New peptide |
ATE553195T1 (en) | 2007-02-08 | 2012-04-15 | Basf Plant Science Gmbh | COMPOSITIONS AND METHODS WITH RNA INTERFERENCE OPR3-LIKE GENE FOR CONTROL OF NEMATODES |
CN101631868B (en) | 2007-02-16 | 2016-02-10 | 巴斯福植物科学有限公司 | For regulating the nucleotide sequence of embryo-specific expression in monocotyledons |
WO2008103643A1 (en) | 2007-02-20 | 2008-08-28 | Monsanto Technology, Llc | Invertebrate micrornas |
US7838729B2 (en) | 2007-02-26 | 2010-11-23 | Monsanto Technology Llc | Chloroplast transit peptides for efficient targeting of DMO and uses thereof |
WO2008108668A1 (en) * | 2007-03-08 | 2008-09-12 | The New Zealand Institute For Plant And Food Research Limited | Transferases, epimerases, polynucleotides encoding these and uses thereof |
BRPI0808665B1 (en) | 2007-03-09 | 2022-05-10 | Monsanto Technology Llc | Methods and constructs for producing transgenic soybean, cotton or canola plants using spectinomycin selection |
KR20080084528A (en) | 2007-03-15 | 2008-09-19 | 제네렉스 바이오테라퓨틱스 인크. | Oncolytic vaccinia virus cancer therapy |
DE112008000747T5 (en) | 2007-03-23 | 2010-01-28 | Basf Plant Science Gmbh | Transgenic plants with increased stress tolerance and increased yield |
US7642436B2 (en) | 2007-04-13 | 2010-01-05 | Ball Horticultural Company | Petunia mutant allele |
CA2584934A1 (en) | 2007-04-17 | 2008-10-17 | University Of Guelph | Nitrogen-regulated sugar sensing gene and protein and modulation thereof |
US8609936B2 (en) | 2007-04-27 | 2013-12-17 | Monsanto Technology Llc | Hemipteran-and coleopteran active toxin proteins from Bacillus thuringiensis |
BRPI0811531A2 (en) | 2007-05-04 | 2014-10-07 | Basf Plant Science Gmbh | POLYNUCLEOTIDE, VECTOR, METHOD FOR MANUFACTURING A PIRUVATO KINASE POLYPEPTIDE, MOUNTED PIRUVATO KINASE POLYPEPTIDE, ANTIBODY, TRANSGENIC ORGANISM FOR A HUMAN FEMAIN WITHOUT A HUMAN FEMALE, A HYDROAUS WITHOUT A HUMAN FEMALE PLANT, USE OF TRANSGENIC NON-HUMAN ORGANISM OR SEED, AND NUCLEIC ACID MOLECULA. |
US8278505B2 (en) | 2007-05-09 | 2012-10-02 | Dow Agrosciences, Llc. | Herbicide resistance genes for resistance to aryloxyalkanoate herbicides |
AU2008252996B2 (en) | 2007-05-22 | 2014-01-30 | Basf Plant Science Gmbh | Plants with increased tolerance and/or resistance to environmental stress and increased biomass production |
AR066642A1 (en) | 2007-05-22 | 2009-09-02 | Basf Plant Gmbh | VEGETABLE CELLS AND PLANTS WITH HIGHER TOLERANCE AND / OR RESISTANCE TO ENVIRONMENTAL STRESS AND HIGHER PRODUCTION OF BIOMASS PRODUCED BY SILENCING OF GENES |
MX2009012596A (en) | 2007-05-24 | 2009-12-08 | Du Pont | Dgat genes from yarrowia lipolytica for increased seed storage lipid production and altered fatty acid profiles in soybean. |
BRPI0812060A2 (en) | 2007-05-29 | 2014-10-07 | Basf Plant Science Gmbh | TRANSGENIC PLANT, INSULATED POLYNUCLEOTIDE, ISOLATED POLYPEPTIDE, AND METHODS FOR PRODUCING A TRANSGENIC PLANT AND FOR INCREASING PLANT GROWTH AND / OR INCREASE UNDER NORMAL OR WATER GROWTH AND / OR INCREASING PLANTS |
EP3567113A1 (en) | 2007-06-06 | 2019-11-13 | Monsanto Technology LLC | Genes and uses for plant enhancement |
AR066922A1 (en) | 2007-06-08 | 2009-09-23 | Monsanto Technology Llc | METHODS OF MOLECULAR IMPROVEMENT OF THE GERMOPLASMA OF A PLANT BY DIRECTED SEQUENCING |
US8945653B2 (en) * | 2007-06-21 | 2015-02-03 | Suntava, Llc | Extracted whole corn kernels and improved processed and processable corn produced thereby |
US8153866B2 (en) * | 2007-06-21 | 2012-04-10 | Suntava Llc | Corn inbreds like FAR045 and hybrids thereof |
US20110265221A1 (en) | 2007-07-10 | 2011-10-27 | Monsanto Technology Llc | Transgenic plants with enhanced agronomic traits |
EP2520655A3 (en) | 2007-07-13 | 2012-12-26 | BASF Plant Science GmbH | Transgenic plants with increased stress tolerance and yield |
US20100242134A1 (en) * | 2007-07-26 | 2010-09-23 | Sathish Puthigae | Methods and polynucleotides for improving plants |
AU2008281715A1 (en) | 2007-08-02 | 2009-02-05 | Basf Plant Science Gmbh | Transgenic plants with increased stress tolerance and yield |
EP2173883A1 (en) | 2007-08-03 | 2010-04-14 | Pioneer Hi-Bred International Inc. | Msca1 nucleotide sequences impacting plant male fertility and method of using same |
BRPI0815841A2 (en) | 2007-08-29 | 2015-09-01 | Monsanto Technology Llc | Methods and compositions for improving preferred traits. |
WO2009029831A1 (en) | 2007-08-31 | 2009-03-05 | University Of Chicago | Methods and compositions related to immunizing against staphylococcal lung diseases and conditions |
US8664475B2 (en) | 2007-09-18 | 2014-03-04 | Basf Plant Science Gmbh | Plants with increased yield |
EP2594646A3 (en) | 2007-09-21 | 2013-11-06 | BASF Plant Science GmbH | Plant with increased yield |
CA2699769C (en) | 2007-09-27 | 2020-08-18 | Manju Gupta | Engineered zinc finger proteins targeting 5-enolpyruvyl shikimate-3-phosphate synthase genes |
JP5550041B2 (en) | 2007-09-27 | 2014-07-16 | 塩野義製薬株式会社 | Method for producing adamantane hydroxide using cytochrome P450 |
US20120047586A9 (en) | 2007-10-24 | 2012-02-23 | Otsuka Chemical Co., Ltd | Polypeptide having enhanced effector function |
AU2008319517B2 (en) * | 2007-10-31 | 2013-11-07 | The New Zealand Institute For Plant And Food Research Limited | Resistance gene and uses thereof |
US8097712B2 (en) | 2007-11-07 | 2012-01-17 | Beelogics Inc. | Compositions for conferring tolerance to viral disease in social insects, and the use thereof |
CN101932712B (en) | 2007-11-20 | 2014-05-14 | 先锋国际良种公司 | Maize ethylene signaling genes and modulation of same for improved stress tolerance in plants |
WO2009068588A2 (en) | 2007-11-27 | 2009-06-04 | Basf Plant Science Gmbh | Transgenic plants with increased stress tolerance and yield |
AU2008331564B2 (en) | 2007-12-03 | 2014-01-09 | Agrivida, Inc. | Engineering enzymatically susceptible proteins |
WO2009077611A2 (en) * | 2007-12-19 | 2009-06-25 | Basf Plant Science Gmbh | Plants with increased yield and/or increased tolerance to environmental stress (iy-bm) |
US20100293665A1 (en) * | 2007-12-21 | 2010-11-18 | Basf Plant Science Gmbh | Plants With Increased Yield (KO NUE) |
WO2009082208A2 (en) | 2007-12-21 | 2009-07-02 | Keygene N.V. | Trichome specific promoters |
NZ564691A (en) * | 2007-12-21 | 2010-03-26 | Nz Inst Plant & Food Res Ltd | Glycosyltransferases, polynucleotides encoding these and methods of use |
US8367899B2 (en) | 2007-12-31 | 2013-02-05 | E I Du Pont De Neumours And Company | Gray leaf spot tolerant maize and methods of production |
JP5677703B2 (en) | 2008-01-10 | 2015-02-25 | リサーチ ディベロップメント ファウンデーション | Vaccine and diagnosis for Ehrlichia chaffiensis |
CN102016048A (en) * | 2008-02-27 | 2011-04-13 | 巴斯夫植物科学有限公司 | Plants with increased yield |
WO2009146015A2 (en) * | 2008-03-31 | 2009-12-03 | Ceres, Inc. | Promoter, promoter control elements, and combinations, and uses thereof |
NZ587653A (en) * | 2008-04-03 | 2011-10-28 | Satish Puthigae | Promoter fragment isolated from Lolium perenne |
EP2607488B1 (en) | 2008-04-07 | 2016-11-02 | Monsanto Technology LLC | Plant regulatory elements and uses thereof |
EP2336332A3 (en) | 2008-04-29 | 2011-11-09 | Monsanto Technology LLC | Genes and uses for plant enhancement |
NZ568190A (en) | 2008-05-12 | 2010-09-30 | Nz Inst Plant & Food Res Ltd | Chimeric compositions and methods for regulating plant gene expression |
EP2119786A1 (en) | 2008-05-13 | 2009-11-18 | Expressive Research B.V. | Increased production of health-promoting compounds in plants |
EP2620500A3 (en) | 2008-05-23 | 2014-01-08 | E. I. du Pont de Nemours and Company | DGAT genes from oleaginous organisms for increased seed storage lipid production and altered fatty acid profiles in oilseed plants |
PL2294198T3 (en) * | 2008-05-28 | 2016-07-29 | Insight Genomics Ltd | Methods and compositions for plant improvement |
MX2010013248A (en) * | 2008-06-03 | 2011-01-21 | Vialactia Biosciences Ltd | Compositions and methods for improving plants. |
DK2297307T3 (en) | 2008-06-04 | 2016-07-25 | Cellular Dynamics Int Inc | PROCEDURES FOR THE MANUFACTURE OF IPS CELLS USING NON-VIRAL METHODS |
WO2010002984A1 (en) | 2008-07-01 | 2010-01-07 | Monsanto Technology, Llc | Recombinant dna constructs and methods for modulating expression of a target gene |
WO2010009353A1 (en) | 2008-07-16 | 2010-01-21 | Monsanto Technology Llc | Methods and vectors for producing transgenic plants |
US8419145B2 (en) * | 2008-07-25 | 2013-04-16 | Eastman Kodak Company | Inkjet printhead and method of printing with multiple drop volumes |
EP3330371A1 (en) | 2008-08-12 | 2018-06-06 | Cellular Dynamics International, Inc. | Methods for the production of ips cells |
JP5662149B2 (en) | 2008-08-13 | 2015-01-28 | 協和発酵キリン株式会社 | Genetically modified protein S composition |
WO2010022089A2 (en) | 2008-08-18 | 2010-02-25 | University Of Maryland, Baltimore | Derivatives of apf and methods of use |
DE112009001994T5 (en) | 2008-08-19 | 2012-01-19 | Basf Plant Science Gmbh | Increased yield plants by increasing or generating one or more activities in a plant or part thereof |
EP2329027A1 (en) | 2008-08-20 | 2011-06-08 | BASF Plant Science GmbH | Transgenic plants comprising as transgene a phosphatidate cytidylyltransferase |
NZ570886A (en) | 2008-08-29 | 2011-05-27 | Nz Inst Plant & Food Res Ltd | Method and compositions to reduce polygalacturonase expression in plants for increasing storage-life of fruit |
DE112009002267T5 (en) | 2008-09-23 | 2012-01-19 | Basf Plant Science Gmbh | Plants with increased yield (LT) |
DE112009002213T5 (en) | 2008-09-23 | 2011-07-28 | BASF Plant Science GmbH, 67063 | Transgenic plants with increased yield |
JP5665081B2 (en) | 2008-10-09 | 2015-02-04 | 協和メデックス株式会社 | Novel fructosyl peptide oxidase |
BRPI0924536A2 (en) | 2008-10-23 | 2015-08-11 | Basf Plant Science Gmbh | Method for producing a transgenic cell with increased gamma-aminobutyric acid (gaba) content to increase yield, isolated nucleic acid molecule, nucleic acid construct, vector, host cell, process for producing a polypeptide, polypeptide, antibody, and , cell nucleus, cell, plant cell nucleus, plant cell, plant tissue, propagation material, pollen, progeny, material collected or a plant |
EP2350289A1 (en) | 2008-10-23 | 2011-08-03 | BASF Plant Science GmbH | Plants with increased yield (nue) |
MX2011003616A (en) | 2008-10-30 | 2011-08-15 | Pioneer Hi Bred Int | Manipulation of glutamine synthetases (gs) to improve nitrogen use efficiency and grain yield in higher plants. |
WO2010064934A1 (en) | 2008-12-01 | 2010-06-10 | Vialactia Biosciences (Nz) Limited | Methods and compositions for the improvement of plant tolerance to environmental stresses |
WO2010068738A1 (en) | 2008-12-10 | 2010-06-17 | Dana-Farber Cancer Institute, Inc. | Mek mutations conferring resistance to mek inhibitors |
EP2370575B1 (en) | 2008-12-17 | 2017-11-01 | Dow AgroSciences LLC | Targeted integration into the zp15 locus |
WO2010071418A1 (en) | 2008-12-19 | 2010-06-24 | Keygene N.V. | Recql5 in homologous recombination |
US20110258735A1 (en) | 2008-12-22 | 2011-10-20 | Marie Coffin | Genes and uses for plant enhancement |
US7868688B2 (en) * | 2008-12-30 | 2011-01-11 | Cosmic Circuits Private Limited | Leakage independent very low bandwith current filter |
EP2206723A1 (en) | 2009-01-12 | 2010-07-14 | Bonas, Ulla | Modular DNA-binding domains |
US20110239315A1 (en) | 2009-01-12 | 2011-09-29 | Ulla Bonas | Modular dna-binding domains and methods of use |
JP2012515532A (en) | 2009-01-20 | 2012-07-12 | ラモット アット テル アビブ ユニバーシティ, リミテッド | MIR-21 promoter driven target cancer treatment |
US8269068B2 (en) | 2009-01-22 | 2012-09-18 | Syngenta Participations Ag | Mutant hydroxyphenylpyruvate dioxygenase polypeptides and methods of use |
US9347046B2 (en) | 2009-01-22 | 2016-05-24 | Syngenta Participations Ag | Hydroxyphenylpyruvate dioxygenase polypeptides and methods of use |
CA2750456A1 (en) | 2009-01-28 | 2010-08-05 | Basf Plant Science Company Gmbh | Transgenic plants having altered nitrogen metabolism |
CA2749708A1 (en) | 2009-01-28 | 2010-08-05 | Basf Plant Science Company Gmbh | Engineering nf-yb transcription factors for enhanced drought resistance and increased yield in transgenic plants |
US8507754B2 (en) | 2009-01-31 | 2013-08-13 | University Of North Texas | Engineering lipids in vegetative tissues of plants |
CN102307994A (en) | 2009-02-06 | 2012-01-04 | 先正达参股股份有限公司 | Modification of multidomain enzyme for expression in plants |
WO2010096510A2 (en) | 2009-02-17 | 2010-08-26 | Edenspace Systems Corporation | Tempering of cellulosic biomass |
US8841516B2 (en) | 2009-02-27 | 2014-09-23 | Monsanto Invest B.V. | Fusarium resistant cucumber plants |
WO2010099431A2 (en) | 2009-02-27 | 2010-09-02 | Monsanto Technology Llc | Hydroponic apparatus and methods of use |
US8716553B2 (en) | 2009-03-02 | 2014-05-06 | Pioneer Hi Bred International Inc | NAC transcriptional activators involved in abiotic stress tolerance |
AU2010221135A1 (en) | 2009-03-05 | 2011-09-29 | Metabolix, Inc. | Propagation of transgenic plants |
WO2010102293A1 (en) | 2009-03-06 | 2010-09-10 | Metabolix, Inc. | Method of positive plant selection using sorbitol dehydrogenase |
CA3012998C (en) | 2009-03-19 | 2021-09-07 | Dsm Ip Assets B.V. | Polyunsaturated fatty acid synthase nucleic acid molecules and polypeptides, compositions, and methods of making and uses thereof |
EP2411524A1 (en) | 2009-03-23 | 2012-02-01 | BASF Plant Science Company GmbH | Transgenic plants with altered redox mechanisms and increased yield |
AU2010232014B8 (en) | 2009-04-01 | 2014-03-06 | Insight Genomics Limited | Control of gene expression in plants using a perennial ryegrass (Lolium perenne L.) derived promoter |
WO2011127032A1 (en) | 2010-04-05 | 2011-10-13 | University Of Chicago | Compositions and methods related to protein a (spa) antibodies as an enhancer of immune response |
CN102612523B (en) | 2009-04-03 | 2016-01-20 | 芝加哥大学 | The composition relevant to albumin A (SPA) variant and method |
US8987553B2 (en) | 2009-04-14 | 2015-03-24 | Pioneer Hi Bred International Inc | Modulation of ACC synthase improves plant yield under low nitrogen conditions |
US8071864B2 (en) * | 2009-04-15 | 2011-12-06 | Monsanto Technology Llc | Plants and seeds of corn variety CV897903 |
US8071865B2 (en) * | 2009-04-15 | 2011-12-06 | Monsanto Technology Llc | Plants and seeds of corn variety CV589782 |
US8362332B2 (en) * | 2009-04-15 | 2013-01-29 | Monsanto Technology Llc | Plants and seeds of corn variety CV165560 |
ES2532145T3 (en) | 2009-04-17 | 2015-03-24 | Dow Agrosciences Llc | Cry toxins insecticides DIG-3 |
US8455716B2 (en) | 2009-04-20 | 2013-06-04 | Monsanto Technology Llc | Multiple virus resistance in plants |
EP2424885B1 (en) | 2009-04-28 | 2016-03-23 | Vanderbilt University | Compositions and methods for the treatment of disorders involving epithelial cell apoptosis |
WO2010138971A1 (en) | 2009-05-29 | 2010-12-02 | Edenspace Systems Corporation | Plant gene regulatory elements |
CN105925535A (en) | 2009-06-05 | 2016-09-07 | 细胞动力国际有限公司 | Method Of Reprogramming T Cells And Hematopoietic Cells |
EP2437592A4 (en) | 2009-06-05 | 2013-02-27 | Univ Florida | Isolation and targeted suppression of lignin biosynthetic genes from sugarcane |
MX336182B (en) | 2009-06-08 | 2016-01-11 | Nunhems Bv | Drought tolerant plants. |
MA33436B1 (en) | 2009-06-17 | 2012-07-03 | Monsanto Invest Nv | NEW TOMATO PLANTS |
US8569582B2 (en) * | 2009-06-22 | 2013-10-29 | The Samuel Roberts Noble Foundation | Method for transformation of grasses |
EA201270107A1 (en) | 2009-07-01 | 2012-07-30 | Байер Байосайенс Н.В. | METHODS AND MEANS FOR OBTAINING PLANTS WITH IMPROVED RESISTANCE TO GLYPHOSATE |
AU2010275363A1 (en) | 2009-07-23 | 2012-02-02 | Basf Plant Science Company Gmbh | Plants with increased yield |
WO2011011273A1 (en) | 2009-07-24 | 2011-01-27 | Pioneer Hi-Bred International, Inc. | The use of dimerization domain component stacks to modulate plant architecture |
US20110035843A1 (en) | 2009-08-05 | 2011-02-10 | Pioneer Hi-Bred International, Inc. | Novel eto1 genes and use of same for reduced ethylene and improved stress tolerance in plants |
US20120142532A1 (en) | 2009-08-10 | 2012-06-07 | Monsanto Technology Llc | Low volatility auxin herbicide formulations |
NZ598053A (en) | 2009-08-11 | 2014-01-31 | Sangamo Biosciences Inc | Organisms homozygous for targeted modification |
CA2770854A1 (en) | 2009-08-20 | 2011-02-24 | Pioneer Hi-Bred International, Inc. | Functional expression of shuffled yeast nitrate transporter (ynt1) in maize to improve nitrate uptake under low nitrate environment |
IN2012DN01315A (en) | 2009-08-20 | 2015-06-05 | Pioneer Hi Bred Int | |
RU2692924C2 (en) | 2009-08-31 | 2019-06-28 | Басф Плант Сайенс Компани Гмбх | Regulatory nucleic acid molecules for enhancing seed-specific and/or seed-preferential gene expression in plants |
SG178389A1 (en) | 2009-08-31 | 2012-03-29 | Basf Plant Science Co Gmbh | Regulatory nucleic acid molecules for enhancing constitutive gene expression in plants |
EP3549435A1 (en) | 2009-09-04 | 2019-10-09 | Monsanto Invest B.V. | Tomato plants exhibiting tolerance to continuous light |
WO2011032180A1 (en) | 2009-09-14 | 2011-03-17 | Jennerex, Inc. | Oncolytic vaccinia virus combination cancer therapy |
US8440891B2 (en) | 2009-09-22 | 2013-05-14 | Board of Trustees of the University of Akransas, N.A. | Rice cultivar CL 142-AR |
MX2012003972A (en) | 2009-10-02 | 2012-05-08 | Pionner Hi Bred International Inc | Down-regulation of acc synthase for improved plant performance. |
US8962584B2 (en) | 2009-10-14 | 2015-02-24 | Yissum Research Development Company Of The Hebrew University Of Jerusalem, Ltd. | Compositions for controlling Varroa mites in bees |
US8440892B2 (en) | 2009-10-15 | 2013-05-14 | Board Of Trustees Of The University Of Arkansas, N.A. | Rice cultivar CL 181-AR |
ES2657067T3 (en) | 2009-10-22 | 2018-03-01 | Dow Agrosciences Llc | Genetically modified zinc finger proteins that target plant genes involved in fatty acid biosynthesis |
US8937214B2 (en) | 2009-10-23 | 2015-01-20 | Monsanto Technology Llc | Methods and compositions for expression of transgenes in plants |
EP2314704A1 (en) | 2009-10-26 | 2011-04-27 | Syngenta Participations AG | Tomato fruit having increased firmness |
CA2780707A1 (en) | 2009-11-17 | 2011-05-26 | Basf Plant Science Company Gmbh | Plants with increased yield |
CA2780527C (en) | 2009-11-23 | 2020-12-01 | E. I. Du Pont De Nemours And Company | Sucrose transporter genes for increasing plant seed lipids |
CN102858959B (en) | 2009-12-10 | 2016-02-24 | 渥太华医院研究院 | Oncolytic rhabdovirus |
JP2013513389A (en) | 2009-12-10 | 2013-04-22 | リージェンツ オブ ザ ユニバーシティ オブ ミネソタ | DNA modification mediated by TAL effectors |
US20130055471A1 (en) | 2009-12-15 | 2013-02-28 | Edwin Henricus Antonius HOLMAN | Transgenic Ozone-Resistant Plants |
WO2011076892A1 (en) | 2009-12-23 | 2011-06-30 | Bayer Cropscience Ag | Plants tolerant to hppd inhibitor herbicides |
AR079972A1 (en) | 2009-12-23 | 2012-03-07 | Bayer Cropscience Ag | TOLERANT PLANTS TO INHIBITING HERBICIDES OF HPPD |
AU2010334815B2 (en) | 2009-12-23 | 2015-07-09 | Bayer Intellectual Property Gmbh | Plants tolerant to HPPD inhibitor herbicides |
AR079882A1 (en) | 2009-12-23 | 2012-02-29 | Bayer Cropscience Ag | TOLERANT PLANTS TO INHIBITING HERBICIDES OF HPPD |
EP2516631B1 (en) | 2009-12-23 | 2018-02-14 | Bayer Intellectual Property GmbH | Plants tolerant to hppd inhibitor herbicides |
CA2784148A1 (en) | 2009-12-28 | 2011-07-28 | Pioneer Hi-Bred International, Inc. | Sorghum fertility restorer genotypes and methods of marker-assisted selection |
CN102906267A (en) | 2010-01-06 | 2013-01-30 | 先锋国际良种公司 | Identification of diurnal rhythms in photosynthetic and non-photosynthetic tissues from zea mays and use in improving crop plants |
AR079909A1 (en) | 2010-01-14 | 2012-02-29 | Monsanto Technology Llc | ELEMENTS OF VEGETABLE REGULATION AND ITS USES |
CA2787594C (en) | 2010-01-22 | 2019-02-26 | Dow Agrosciences Llc | Engineered landing pads for gene targeting in plants |
KR101951489B1 (en) | 2010-01-22 | 2019-04-22 | 다우 아그로사이언시즈 엘엘씨 | Excision of transgenes in genetically modified organisms |
AR080021A1 (en) | 2010-01-26 | 2012-03-07 | Pioneer Hi Bred Int | TOLERANCE TO HPPD INHIBITING HERBICIDES (HYDROPHENYL PIRUVATO DIOXYGENASE) |
MY169476A (en) | 2010-02-04 | 2019-04-12 | Bayer Cropscience Ag | A method for increasing photosynthetic carbon fixation using glycolate dehydrogenase multi-subunit fusion protein |
EP2660318A1 (en) | 2010-02-09 | 2013-11-06 | Sangamo BioSciences, Inc. | Targeted genomic modification with partially single-stranded donor molecules |
US10080799B2 (en) | 2010-02-12 | 2018-09-25 | Arizona Board Of Regents On Behalf Of Arizona State University | Methods and compositions related to glycoprotein-immunoglobulin fusions |
EP2538943B1 (en) | 2010-02-25 | 2016-03-30 | Dana-Farber Cancer Institute, Inc. | Braf mutations conferring resistance to braf inhibitors |
WO2011108502A1 (en) | 2010-03-02 | 2011-09-09 | 協和発酵キリン株式会社 | Modified antibody composition |
BR112012022218A2 (en) | 2010-03-04 | 2018-07-10 | Mendel Biotechnology Inc | transcription regulators to improve plant performance |
EP3214174B1 (en) | 2010-03-04 | 2019-10-16 | InteRNA Technologies B.V. | A mirna molecule defined by its source and its diagnostic and therapeutic uses in diseases or conditions associated with emt |
EA201792402A3 (en) | 2010-03-08 | 2018-09-28 | Монсанто Текнолоджи Ллс | POLYNUCLEOTIDE MOLECULES FOR REGULATION OF PLANT GENES |
EP2554669B1 (en) | 2010-03-26 | 2018-09-19 | Kyowa Hakko Kirin Co., Ltd. | Novel antibody having modification site introduced therein, and antibody fragment |
WO2011133512A1 (en) | 2010-04-19 | 2011-10-27 | Research Development Foundation | Rtef-1 variants and uses thereof |
EP2380905A1 (en) | 2010-04-19 | 2011-10-26 | Thrombotargets Europe, S.L. | Phospholipid-enriched vesicles bearing tissue factor having haemostatic activities and uses thereof |
BR112012027208A2 (en) | 2010-04-23 | 2015-09-15 | Dow Agrosciences Llc | combinations including cry3aa and cry6aa proteins to prevent the development of resistance in maize root system chrysomelids (diabrotica spp). |
US9234208B1 (en) | 2010-05-10 | 2016-01-12 | Dow Agrosciences Llc | DIG-13 insecticidal cry toxins |
EP2571512B1 (en) | 2010-05-17 | 2017-08-23 | Sangamo BioSciences, Inc. | Novel dna-binding proteins and uses thereof |
AU2011255481B2 (en) | 2010-05-19 | 2014-09-11 | The Samuel Roberts Noble Foundation, Inc. | Altered leaf morphology and enhanced agronomic properties in plants |
US20130145499A1 (en) | 2010-05-28 | 2013-06-06 | Willem Hendrik Vriezen | Plants with Increased Fruit Size |
MX342966B (en) | 2010-06-09 | 2016-10-19 | Dana Farber Cancer Inst Inc | A mek 1 mutation conferring resistance to raf and mek inhibitors. |
KR101861168B1 (en) | 2010-06-15 | 2018-05-31 | 후지필름 셀룰러 다이내믹스, 인코포레이티드 | Generation of induced pluripotent stem cells from small volumes of peripheral blood |
JP2013530699A (en) | 2010-06-15 | 2013-08-01 | セルラー ダイナミクス インターナショナル, インコーポレイテッド | Overview of ready-made stem cell models for investigating biological responses |
FR2961375B1 (en) | 2010-06-16 | 2016-05-13 | Inst De Rech Pour Le Dev (Ird) | OVERPRODUCTION OF JASMONIC ACID IN TRANSGENIC PLANTS |
ES2623454T3 (en) | 2010-06-24 | 2017-07-11 | Dow Agrosciences Llc | Reduction of the content of saturated fatty acids in plant seeds |
AR081302A1 (en) | 2010-06-24 | 2012-08-01 | Brookhaven Science Ass Llc | ACCUMULATION OF OMEGA-7 FATTY ACIDS (-7) IN PLANT SEEDS |
WO2011163292A1 (en) * | 2010-06-24 | 2011-12-29 | Syngenta Participations Ag | Methods for agrobacterium-mediated transformation of sugar cane |
CA2803298C (en) | 2010-07-02 | 2020-07-14 | The University Of Chicago | Compositions and methods related to protein a (spa) variants |
EP3369817A1 (en) | 2010-07-06 | 2018-09-05 | InteRNA Technologies B.V. | Mirna and its diagnostic and therapeutic uses in diseases or conditions associated with melanoma , or in diseases or conditions with activated braf pathway |
JP5897002B2 (en) | 2010-07-07 | 2016-04-13 | セルラー ダイナミクス インターナショナル, インコーポレイテッド | Endothelial cell production by programming |
JP5058332B2 (en) | 2010-07-14 | 2012-10-24 | 住友ゴム工業株式会社 | Isoprene oligomer, polyisoprene, and production methods thereof, rubber composition, and pneumatic tire |
WO2012012741A1 (en) | 2010-07-23 | 2012-01-26 | Board Of Trustees Of Michigan State University | FERULOYL-CoA:MONOLIGNOL TRANSFERASE |
US9617551B2 (en) | 2010-07-29 | 2017-04-11 | Dow Agrosciences Llc | Method of increasing plant transformation frequency using modified strains of Agrobacteria |
EP2601289B1 (en) | 2010-08-04 | 2017-07-12 | Cellular Dynamics International, Inc. | Reprogramming immortalized b cells |
US8816153B2 (en) | 2010-08-27 | 2014-08-26 | Monsanto Technology Llc | Recombinant DNA constructs employing site-specific recombination |
EP2611924B1 (en) | 2010-08-30 | 2019-03-06 | Dow Agrosciences LLC | Activation tagging platform for maize, and resultant tagged population and plants |
JP6039560B2 (en) | 2010-08-30 | 2016-12-07 | ダウ アグロサイエンシィズ エルエルシー | Sugarcane-boiled virus (SCBV) enhancer and its use in plant functional genomics |
US9095540B2 (en) | 2010-09-09 | 2015-08-04 | The University Of Chicago | Methods and compositions involving protective staphylococcal antigens |
US9045549B2 (en) | 2010-11-03 | 2015-06-02 | The Samuel Roberts Noble Foundation, Inc. | Transcription factors for modification of lignin content in plants |
WO2012074868A2 (en) | 2010-12-03 | 2012-06-07 | Ms Technologies, Llc | Optimized expression of glyphosate resistance encoding nucleic acid molecules in plant cells |
GB201020737D0 (en) | 2010-12-08 | 2011-01-19 | Univ Leuven Kath | Anti-oxidative stress agents |
EP2651207B1 (en) | 2010-12-17 | 2017-11-08 | Monsanto Technology LLC | Methods for improving competency of plant cells |
CA2823467C (en) | 2010-12-22 | 2020-10-27 | Pioneer Hi-Bred International, Inc. | Qtls associated with and methods for identifying whole plant field resistance to sclerotinia |
CN106978439A (en) | 2010-12-30 | 2017-07-25 | 陶氏益农公司 | Assign the nucleic acid molecules to the resistance of coleoptera harmful organism |
US8987552B2 (en) | 2010-12-30 | 2015-03-24 | Dow Agrosciences, Llc. | Nucleic acid molecules that target the vacuolar ATPase H subunit and confer resistance to coleopteran pests |
CN105779450A (en) | 2010-12-30 | 2016-07-20 | 陶氏益农公司 | Nucleic acid molecules that target the vacuolar ATPase C subunit and confer resistance to coleopteran pests |
AU2012204467B2 (en) | 2011-01-04 | 2016-08-18 | Sillajen, Inc. | Generation of antibodies to tumor antigens and generation of tumor specific complement dependent cytotoxicity by administration of oncolytic vaccinia virus |
WO2012095841A1 (en) | 2011-01-10 | 2012-07-19 | State Of Israel, Ministry Of Agriculture And Rural Development, A.R.O. - Volcani Center | Improved tomato plants |
EP2474617A1 (en) | 2011-01-11 | 2012-07-11 | InteRNA Technologies BV | Mir for treating neo-angiogenesis |
JP6005666B2 (en) | 2011-02-08 | 2016-10-12 | セルラー ダイナミクス インターナショナル, インコーポレイテッド | Production of hematopoietic progenitor cells by programming |
US8648230B2 (en) | 2011-03-18 | 2014-02-11 | Ms Technologies, Llc | Regulatory regions preferentially expressing in non-pollen plant tissue |
EP3406628A1 (en) | 2011-04-08 | 2018-11-28 | Evaxion Biotech ApS | Proteins and nucleic acids useful in vaccines targeting staphylococcus aureus |
US9556448B2 (en) | 2011-04-11 | 2017-01-31 | Targeted Growth, Inc. | Identification and the use of KRP mutants in plants |
UY34014A (en) | 2011-04-15 | 2012-11-30 | Dow Agrosciences Llc | SYNTHETIC GENES TO EXPRESS PROTEINS IN CORN CELLS, CONSTRUCTIONS, TRANSGENIC PLANTS, PEST CONTROL METHODS AND COMPOSITIONS |
EP2514303A1 (en) | 2011-04-21 | 2012-10-24 | Agventure B.V. | Hybrid seed potato breeding |
EP2794643A1 (en) | 2011-04-29 | 2014-10-29 | Pioneer Hi-Bred International Inc. | Down-regulation of a homeodomain-leucine zipper i-class homeobox gene for improved plant performance |
EP2702157A1 (en) | 2011-04-29 | 2014-03-05 | Keygene N.V. | Glyphosate resistance enhancement |
US8901371B2 (en) | 2011-05-06 | 2014-12-02 | The Samuel Roberts Noble Foundation, Inc. | Compositions and methods for improved plant feedstock |
US8945588B2 (en) | 2011-05-06 | 2015-02-03 | The University Of Chicago | Methods and compositions involving protective staphylococcal antigens, such as EBH polypeptides |
WO2012159891A1 (en) | 2011-05-20 | 2012-11-29 | Syngenta Participations Ag | Endosperm-specific plant promoters and uses therefor |
EP2714901A1 (en) | 2011-05-31 | 2014-04-09 | Keygene N.V. | Pest resistant plants |
EP2718443B1 (en) | 2011-06-06 | 2017-11-29 | Bayer CropScience NV | Methods and means to modify a plant genome at a preselected site |
ES2627529T3 (en) | 2011-06-08 | 2017-07-28 | Children's Hospital Of Eastern Ontario Research Institute Inc. | Compositions for glioblastoma treatment |
WO2013006861A1 (en) | 2011-07-07 | 2013-01-10 | University Of Georgia Research Foundation, Inc. | Sorghum grain shattering gene and uses thereof in altering seed dispersal |
EP2737067A1 (en) | 2011-07-28 | 2014-06-04 | Genective | Glyphosate tolerant corn event vco-ø1981-5 and kit and method for detecting the same |
AU2012296576B2 (en) | 2011-08-15 | 2017-09-07 | The University Of Chicago | Compositions and methods related to antibodies to staphylococcal protein a |
MX348003B (en) | 2011-08-22 | 2017-03-08 | Bayer Cropscience Nv | Methods and means to modify a plant genome. |
US11377663B1 (en) | 2011-08-30 | 2022-07-05 | Monsanto Technology Llc | Genetic regulatory elements |
AR088155A1 (en) | 2011-09-13 | 2014-05-14 | Monsanto Technology Llc | METHODS AND COMPOSITIONS FOR WEED CONTROL |
AU2012308765B2 (en) | 2011-09-13 | 2018-06-21 | Monsanto Technology Llc | Methods and compositions for weed control |
WO2013040057A1 (en) | 2011-09-13 | 2013-03-21 | Monsanto Technology Llc | Methods and compositions for weed control |
US10806146B2 (en) | 2011-09-13 | 2020-10-20 | Monsanto Technology Llc | Methods and compositions for weed control |
US10829828B2 (en) | 2011-09-13 | 2020-11-10 | Monsanto Technology Llc | Methods and compositions for weed control |
EP2755467B1 (en) | 2011-09-13 | 2017-07-19 | Monsanto Technology LLC | Methods and compositions for weed control |
US10760086B2 (en) | 2011-09-13 | 2020-09-01 | Monsanto Technology Llc | Methods and compositions for weed control |
US9840715B1 (en) | 2011-09-13 | 2017-12-12 | Monsanto Technology Llc | Methods and compositions for delaying senescence and improving disease tolerance and yield in plants |
US9920326B1 (en) | 2011-09-14 | 2018-03-20 | Monsanto Technology Llc | Methods and compositions for increasing invertase activity in plants |
WO2013048255A2 (en) | 2011-09-30 | 2013-04-04 | Keygene N.V. | Heat tolerance microrna |
WO2013048254A1 (en) | 2011-09-30 | 2013-04-04 | Keygene N.V. | Heat tolerance microrna |
AU2012318626B2 (en) | 2011-10-06 | 2017-07-20 | Board Of Trustees Of Michigan State University | Hibiscus cannabinus feruloyl-CoA:monolignol transferase |
US9273102B2 (en) | 2011-10-12 | 2016-03-01 | Niels Iversen Møller | Peptides derived from Campylobacter jejuni and their use in vaccination |
CA2853449A1 (en) | 2011-10-25 | 2013-05-02 | Pioneer Hi-Bred International, Inc. | Methods to alter plant cell wall composition for improved biofuel production and silage digestibility |
UY39628A (en) | 2011-10-26 | 2022-03-31 | Monsanto Technology Llc | HERBICIDE SALTS OF CARBOXYLIC ACID |
BR112014010537A2 (en) | 2011-10-31 | 2017-05-02 | Pioneer Hi Bred Int | method for modulating ethylene sensitivity, transgenic plant, isolated protein, isolated polynucleotide sequence, polypeptide with ethylene regulatory activity, method for increasing yield in a plant, method for improving an agronomic parameter of a plant, method assisted by selection marker of a plant |
FR2984076A1 (en) | 2011-12-15 | 2013-06-21 | Inst Rech Developpement Ird | OVERPRODUCTION OF JASMONATES IN TRANSGENIC PLANTS |
WO2013090814A2 (en) | 2011-12-16 | 2013-06-20 | Board Of Trustees Of Michigan State University | p-Coumaroyl-CoA:Monolignol Transferase |
WO2013095132A1 (en) | 2011-12-22 | 2013-06-27 | Interna Technologies B.V. | Mirna for treating head and neck cancer |
UY34607A (en) | 2012-02-01 | 2013-09-02 | Dow Agrosciences Llc | GLIFOSATO RESISTANT PLANTS AND ASSOCIATED METHODS. |
US10351610B2 (en) | 2012-02-02 | 2019-07-16 | Dow Agrosciences Llc | Plant transactivation interaction motifs and uses thereof |
AU2013214833B2 (en) | 2012-02-02 | 2018-04-19 | Conicet | HaHB11 provides improved plant yield and tolerance to abiotic stress |
CA3204007A1 (en) | 2012-02-27 | 2013-09-06 | Board Of Trustees Of Michigan State University | Control of cellulose biosynthesis by overexpression of a transcription factor myb46 |
CN108070600B (en) | 2012-02-29 | 2021-09-21 | 先正达参股股份有限公司 | Modulation of seed vigor |
WO2013130813A1 (en) | 2012-02-29 | 2013-09-06 | Dow Agrosciences Llc | Sugarcane bacilliform viral (scbv) enhancer and its use in plant functional genomics |
WO2013134651A1 (en) | 2012-03-09 | 2013-09-12 | Board Of Trustees Of Michigan State University | Method of enhancing plant drought tolerance by expression of ndr1 |
WO2013136274A1 (en) | 2012-03-13 | 2013-09-19 | University Of Guelph | Myb55 promoter and use thereof |
MX2014011038A (en) | 2012-03-13 | 2015-06-02 | Pioneer Hi Bred Int | Genetic reduction of male fertility in plants. |
US20150218578A1 (en) | 2012-03-13 | 2015-08-06 | University Of Guelph | Methods of increasing tolerance to heat stress and amino acid content of plants |
WO2013138309A1 (en) | 2012-03-13 | 2013-09-19 | Pioneer Hi-Bred International, Inc. | Genetic reduction of male fertility in plants |
EP2831099B1 (en) | 2012-03-26 | 2020-04-22 | Axcella Health Inc. | Nutritive fragments, proteins and methods |
ES2727932T3 (en) | 2012-04-12 | 2019-10-21 | Syngenta Participations Ag | Qtl responsible for the firmness of tomato fruits |
AU2013205557B2 (en) | 2012-04-17 | 2016-04-21 | Corteva Agriscience Llc | Synthetic brassica-derived chloroplast transit peptides |
US10155959B2 (en) | 2012-04-20 | 2018-12-18 | Futuragene Israel Ltd. | Bronze bug control agents |
CN104508134A (en) | 2012-04-23 | 2015-04-08 | 富优基尼以色列股份有限公司 | Glycaspis brimblecombei control agents |
EP2841581B2 (en) | 2012-04-23 | 2023-03-08 | BASF Agricultural Solutions Seed US LLC | Targeted genome engineering in plants |
WO2013162751A1 (en) | 2012-04-26 | 2013-10-31 | University Of Chicago | Compositions and methods related to antibodies that neutralize coagulase activity during staphylococcus aureus disease |
CA2910319C (en) | 2012-04-26 | 2022-06-14 | University Of Chicago | Staphylococcal coagulase antigens and methods of their use |
MX369788B (en) | 2012-05-02 | 2019-11-21 | Dow Agrosciences Llc | Targeted modification of malate dehydrogenase. |
EP2847338B1 (en) | 2012-05-07 | 2018-09-19 | Sangamo Therapeutics, Inc. | Methods and compositions for nuclease-mediated targeted integration of transgenes |
WO2013173535A2 (en) | 2012-05-18 | 2013-11-21 | E. I. Du Pont De Nemours And Company | Inducible promoter sequences for regulated expression and methods of use |
AR091143A1 (en) | 2012-05-24 | 2015-01-14 | Seeds Ltd Ab | COMPOSITIONS AND METHODS TO SILENCE GENETIC EXPRESSION |
WO2013180809A1 (en) | 2012-05-29 | 2013-12-05 | North Carolina Central University | Methods for the production of cytoprotective asialo-erythropoietin in plants and its purification from plant tissues |
UY34845A (en) | 2012-06-04 | 2014-01-31 | Monsanto Technology Llc | ? WATER CONCENTRATED HERBICIDE COMPOSITIONS CONTAINING GLIFOSATE SALTS AND DICAMBA SALTS |
WO2013184768A1 (en) | 2012-06-05 | 2013-12-12 | University Of Georgia Research Foundation, Inc. | Compositions and methods of gene silencing in plants |
US20150225734A1 (en) | 2012-06-19 | 2015-08-13 | Regents Of The University Of Minnesota | Gene targeting in plants using dna viruses |
NZ629197A (en) | 2012-06-26 | 2016-07-29 | Forage Genetics Internat Llc | Methods and composition for enhanced forage quality |
CA2881787A1 (en) | 2012-08-13 | 2014-02-20 | University Of Georgia Research Foundation, Inc. | Compositions and methods for increasing pest resistance in plants |
AU2013302947B2 (en) | 2012-08-17 | 2017-06-01 | Diaa ALABED | Use of a maize untranslated region for transgene expression in plants |
AR092482A1 (en) | 2012-09-07 | 2015-04-22 | Dow Agrosciences Llc | ENRICHMENT OF THE CLASSIFICATION OF FLUORESCENCE ACTIVATED CELLS (FACS) TO GENERATE PLANTS |
RU2665811C2 (en) | 2012-09-07 | 2018-09-04 | ДАУ АГРОСАЙЕНСИЗ ЭлЭлСи | Fad3 performance loci and corresponding target site specific binding proteins capable of inducing targeted breaks |
UA118090C2 (en) | 2012-09-07 | 2018-11-26 | ДАУ АГРОСАЙЄНСІЗ ЕлЕлСі | Fad2 performance loci and corresponding target site specific binding proteins capable of inducing targeted breaks |
WO2014042923A1 (en) | 2012-09-13 | 2014-03-20 | Indiana University Research & Technology Corporation | Compositions and systems for conferring disease resistance in plants and methods of use thereof |
BR112015005674B1 (en) | 2012-09-14 | 2022-09-06 | BASF Agricultural Solutions Seed US LLC | RECOMBINANT POLYPEPTIDES TO PROVIDE HERBICIDES TOLERANCE |
WO2014047623A1 (en) | 2012-09-24 | 2014-03-27 | Monsanto Technology Llc | Method for improving shelf life by regulating expression of polyphenol oxidase |
US20150259701A1 (en) | 2012-10-03 | 2015-09-17 | Futuragene Israel Ltd. | Gall wasp control agents |
EP2908620A4 (en) | 2012-10-18 | 2016-07-27 | Monsanto Technology Llc | Methods and compositions for plant pest control |
AU2013340443B2 (en) | 2012-10-30 | 2019-06-06 | Agresearch Limited | Enhanced acyltransferase polynucleotides, polypeptides, and methods of use |
US9896694B2 (en) | 2012-10-30 | 2018-02-20 | Agresearch Limited | Acyltransferase polynucleotides, polypeptides and methods of use |
US9957519B2 (en) | 2012-10-30 | 2018-05-01 | Agresearch Limited | Acyltransferase polynucleotides, polypeptides and methods of use |
NZ707560A (en) | 2012-11-01 | 2019-10-25 | Cellectis | Plants for production of therapeutic proteins |
WO2014072357A1 (en) | 2012-11-06 | 2014-05-15 | Interna Technologies B.V. | Combination for use in treating diseases or conditions associated with melanoma, or treating diseases or conditions associated with activated b-raf pathway |
CA2889557A1 (en) | 2012-11-20 | 2014-05-30 | Pioneer Hi-Bred International., Inc. | Engineering plants for efficient uptake and utilization of urea to improve crop production |
US9598707B2 (en) | 2012-11-26 | 2017-03-21 | Arkansas State University-Jonesboro | Method to increase the yield of products in plant material |
WO2014084884A1 (en) | 2012-11-28 | 2014-06-05 | Monsanto Technology Llc | Transgenic plants with enhanced traits |
US10253325B2 (en) | 2012-12-19 | 2019-04-09 | Boston Medical Center Corporation | Methods for elevating fat/oil content in plants |
CA2894979A1 (en) | 2012-12-21 | 2014-06-26 | The New Zealand Institute For Plant And Food Research Limited | Regulation of gene expression |
EP2934097B1 (en) | 2012-12-21 | 2018-05-02 | Cellectis | Potatoes with reduced cold-induced sweetening |
EP2941488B1 (en) | 2013-01-01 | 2023-03-22 | Monsanto Technology LLC | Methods of introducing dsrna to plant seeds for modulating gene expression |
US10683505B2 (en) | 2013-01-01 | 2020-06-16 | Monsanto Technology Llc | Methods of introducing dsRNA to plant seeds for modulating gene expression |
CN113005148A (en) | 2013-01-16 | 2021-06-22 | 爱默蕾大学 | CAS 9-nucleic acid complexes and uses related thereto |
WO2014116721A1 (en) | 2013-01-22 | 2014-07-31 | The Arizona Board Of Regents For And On Behalf Of Arizona State University | Geminiviral vector for expression of rituximab |
US10000767B2 (en) | 2013-01-28 | 2018-06-19 | Monsanto Technology Llc | Methods and compositions for plant pest control |
CN104955324A (en) | 2013-01-29 | 2015-09-30 | 巴斯夫植物科学有限公司 | Fungal resistant plants expressing HCP7 |
BR112015017345A2 (en) | 2013-01-29 | 2017-11-21 | Basf Plant Science Co Gmbh | method for increasing fungal resistance in a plant, recombinant vector construction, transgenic plant, method for producing transgency plant, use of any of the exogenous nucleic acids, part that can be harvested from a transgenic plant, product derived from a plant , method for producing a product and method for growing a fungal resistant plant |
CA2891424A1 (en) | 2013-01-29 | 2014-08-07 | Basf Plant Science Company Gmbh | Fungal resistant plants expressing ein2 |
MX2015010783A (en) | 2013-02-21 | 2016-06-21 | Children S Hospital Of Eastern Ontario Res Inst Inc | Vaccine composition. |
WO2014130770A1 (en) | 2013-02-22 | 2014-08-28 | Cellular Dynamics International, Inc. | Hepatocyte production via forward programming by combined genetic and chemical engineering |
CA2901676C (en) | 2013-02-25 | 2023-08-22 | Sangamo Biosciences, Inc. | Methods and compositions for enhancing nuclease-mediated gene disruption |
TR201815726T4 (en) | 2013-02-27 | 2018-11-21 | Monsanto Technology Llc | Glyphosate and dicamba tank mixtures with improved volatility. |
US20160010094A1 (en) | 2013-03-01 | 2016-01-14 | University Of Pretoria | Transgenic cell selection |
US9783817B2 (en) | 2013-03-04 | 2017-10-10 | Arkansas State University | Methods of expressing and detecting activity of expansin in plant cells |
WO2014135682A1 (en) | 2013-03-08 | 2014-09-12 | Basf Plant Science Company Gmbh | Fungal resistant plants expressing mybtf |
US20160002648A1 (en) | 2013-03-11 | 2016-01-07 | Mei Guo | Genes for improving nutrient uptake and abiotic stress tolerance in plants |
MX364458B (en) | 2013-03-13 | 2019-04-26 | Monsanto Technology Llc | Methods and compositions for weed control. |
US20160017360A1 (en) | 2013-03-13 | 2016-01-21 | Pioneer Hi-Bred International, Inc. | Functional expression of bacterial major facilitator superfamily mfs gene in maize to improve agronomic traits and grain yield |
EP2967082A4 (en) | 2013-03-13 | 2016-11-02 | Monsanto Technology Llc | Methods and compositions for weed control |
US20160010101A1 (en) | 2013-03-13 | 2016-01-14 | Pioneer Hi-Bred International, Inc. | Enhanced nitrate uptake and nitrate translocation by over- expressing maize functional low-affinity nitrate transporters in transgenic maize |
EP2971001A4 (en) | 2013-03-14 | 2016-08-17 | Evolutionary Genomics Inc | Identification and use of tomato genes controlling salt/drought tolerance and fruit sweetness |
US20140283211A1 (en) | 2013-03-14 | 2014-09-18 | Monsanto Technology Llc | Methods and Compositions for Plant Pest Control |
US20140304857A1 (en) | 2013-03-14 | 2014-10-09 | Pioneer Hi Bred International Inc | Maize stress related transcription factor 18 and uses thereof |
US20160024516A1 (en) | 2013-03-15 | 2016-01-28 | Pioneer Hi-Bred International, Inc. | Modulation of ACC Deaminase Expression |
US10568328B2 (en) | 2013-03-15 | 2020-02-25 | Monsanto Technology Llc | Methods and compositions for weed control |
US10113162B2 (en) | 2013-03-15 | 2018-10-30 | Cellectis | Modifying soybean oil composition through targeted knockout of the FAD2-1A/1B genes |
JP6433886B2 (en) * | 2013-03-15 | 2018-12-05 | サントリーホールディングス株式会社 | Plant cell wall treating agent, substance delivery method and substance delivery system using the treating agent |
CN105143454A (en) | 2013-03-15 | 2015-12-09 | 先锋国际良种公司 | Compositions and methods of use of ACC oxidase polynucleotides and polypeptides |
JP2016514465A (en) | 2013-03-22 | 2016-05-23 | フラウンホーファー−ゲゼルシャフト ツール フエルデルング デア アンゲヴァンテン フォルシュング エー.ファオ. | Kits containing plus-sense single-stranded RNA viral vectors and methods for making polypeptides using these kits |
KR102192599B1 (en) | 2013-04-05 | 2020-12-18 | 다우 아그로사이언시즈 엘엘씨 | Methods and compositions for integration of an exogenous sequence within the genome of plants |
US10526378B2 (en) | 2013-04-19 | 2020-01-07 | Agresearch Limited | Methods and materials for encapsulating proteins |
EP3730615A3 (en) | 2013-05-15 | 2020-12-09 | Sangamo Therapeutics, Inc. | Methods and compositions for treatment of a genetic condition |
EP2810952A1 (en) | 2013-06-03 | 2014-12-10 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Novel pest control methods |
WO2015005258A1 (en) | 2013-07-09 | 2015-01-15 | 協和メデックス株式会社 | Glycated hexapeptide oxidase and use thereof |
US9850496B2 (en) | 2013-07-19 | 2017-12-26 | Monsanto Technology Llc | Compositions and methods for controlling Leptinotarsa |
MX359191B (en) | 2013-07-19 | 2018-09-18 | Monsanto Technology Llc | Compositions and methods for controlling leptinotarsa. |
AU2014317861B2 (en) | 2013-09-09 | 2019-11-28 | Figene, Llc | Gene therapy for the regeneration of chondrocytes or cartilage type cells |
JP2016536978A (en) | 2013-10-04 | 2016-12-01 | ダウ アグロサイエンシィズ エルエルシー | Maize (Zea mays) metallothionein-like regulatory elements and uses thereof |
US20160237447A1 (en) | 2013-10-07 | 2016-08-18 | Monsanto Technology Llc | Transgenic Plants With Enhanced Traits |
BR102014025499A2 (en) | 2013-10-15 | 2015-09-29 | Dow Agrosciences Llc | zea mays regulatory elements and their use |
BR102014025574A2 (en) | 2013-10-15 | 2015-09-29 | Dow Agrosciences Llc | zea mays regulatory elements and uses thereof |
CA3013907C (en) | 2013-10-29 | 2020-09-08 | Biotech Institute, Llc | Production and use of specialty cannabis with bd/bt genotype and a limonene-dominant terpene profile |
UA120503C2 (en) | 2013-11-04 | 2019-12-26 | Дау Агросайєнсиз Елелсі | Optimal maize loci |
BR102014027438B1 (en) | 2013-11-04 | 2022-09-27 | Dow Agrosciences Llc | RECOMBINANT NUCLEIC ACID MOLECULE AND PRODUCTION METHOD OF A TRANSGENIC PLANT CELL |
AR098295A1 (en) | 2013-11-04 | 2016-05-26 | Monsanto Technology Llc | COMPOSITIONS AND METHODS TO CONTROL INFESTATIONS OF PESTS AND PARASITES OF THE ARTHROPODS |
JP6560205B2 (en) | 2013-11-04 | 2019-08-14 | ダウ アグロサイエンシィズ エルエルシー | Optimal soybean locus |
US10028902B2 (en) | 2013-11-08 | 2018-07-24 | Baylor Research Institute | Nuclear localization of GLP-1 stimulates myocardial regeneration and reverses heart failure |
TW201525136A (en) | 2013-11-26 | 2015-07-01 | Dow Agrosciences Llc | Production of omega-3 long-chain polyunsaturated fatty acids in oilseed crops by a thraustochytrid PUFA synthase |
EP3077820A1 (en) | 2013-12-03 | 2016-10-12 | Evaxion Biotech ApS | Proteins and nucleic acids useful in vaccines targeting staphylococcus aureus |
UA119253C2 (en) | 2013-12-10 | 2019-05-27 | Біолоджикс, Інк. | Compositions and methods for virus control in varroa mite and bees |
NL2011980C2 (en) | 2013-12-17 | 2015-06-18 | Univ Leiden | New effects of plant ahl proteins. |
BR102014031844A2 (en) | 2013-12-20 | 2015-10-06 | Dow Agrosciences Llc | RAS and related nucleic acid molecules that confer resistance to Coleoptera and Hemiptera pests |
WO2015095750A1 (en) | 2013-12-20 | 2015-06-25 | Dow Agrosciences Llc | Rnapii-140 nucleic acid molecules that confer resistance to coleopteran pests |
TW201527312A (en) | 2013-12-31 | 2015-07-16 | Dow Agrosciences Llc | Novel maize ubiquitin promoters |
TW201527313A (en) | 2013-12-31 | 2015-07-16 | Dow Agrosciences Llc | Novel maize ubiquitin promoters |
US10683513B2 (en) | 2013-12-31 | 2020-06-16 | Dow Agrosciences Llc | Tissue-specific expression and hybrid plant production |
TW201527314A (en) | 2013-12-31 | 2015-07-16 | Dow Agrosciences Llc | Novel maize ubiquitin promoters |
TW201527316A (en) | 2013-12-31 | 2015-07-16 | Dow Agrosciences Llc | Novel maize ubiquitin promoters |
AR099092A1 (en) | 2014-01-15 | 2016-06-29 | Monsanto Technology Llc | METHODS AND COMPOSITIONS FOR WEED CONTROL USING EPSPS POLYUCLEOTIDES |
BR102015000943A2 (en) | 2014-01-17 | 2016-06-07 | Dow Agrosciences Llc | increased protein expression in plant |
JP6448194B2 (en) | 2014-01-20 | 2019-01-09 | 住友ゴム工業株式会社 | By introducing a gene encoding cis-prenyltransferase and a gene encoding NogoBreceptor into the host, the transformant in which the cis-prenyltransferase and NogoBreceptor were expressed in the host, and the transformant were used. Method for producing polyisoprenoid |
US9714429B2 (en) | 2014-01-28 | 2017-07-25 | Arkansas State University | Regulatory sequence of cupin family gene |
EP3105332A4 (en) | 2014-02-14 | 2018-01-10 | University of Utah Research Foundation | Methods and compositions for inhibiting retinopathy of prematurity |
WO2015130783A1 (en) | 2014-02-25 | 2015-09-03 | Research Development Foundation | Sty peptides for inhibition of angiogenesis |
EP3971283A1 (en) | 2014-02-27 | 2022-03-23 | Monsanto Technology LLC | Compositions and methods for site directed genomic modification |
TW201538518A (en) | 2014-02-28 | 2015-10-16 | Dow Agrosciences Llc | Root specific expression conferred by chimeric gene regulatory elements |
BR112016020889B1 (en) | 2014-03-11 | 2022-10-04 | BASF Agricultural Solutions Seed US LLC | RECOMBINANT NUCLEIC ACID MOLECULE, BACTERIAL HOST CELL, RECOMBINANT HPPD PROTEIN, RECOMBINANT NUCLEIC ACID USE AND BASE PRODUCT |
TW201542093A (en) | 2014-03-21 | 2015-11-16 | 艾格里遺傳學股份有限公司 | Cry1D for controlling corn earworm |
US11091770B2 (en) | 2014-04-01 | 2021-08-17 | Monsanto Technology Llc | Compositions and methods for controlling insect pests |
WO2015164228A1 (en) | 2014-04-21 | 2015-10-29 | Cellular Dynamics International, Inc. | Hepatocyte production via forward programming by combined genetic and chemical engineering |
WO2015168124A1 (en) | 2014-04-28 | 2015-11-05 | The Trustees Of The University Of Pennsylvania | Compositions and methods for controlling plant growth and development |
RU2016146483A (en) | 2014-05-07 | 2018-06-09 | ДАУ АГРОСАЙЕНСИЗ ЭлЭлСи | DRE4 NUCLEIC ACID MOLECULES COMMUNICATING RESISTANCE TO RACID PEST PEST |
WO2015183096A1 (en) | 2014-05-30 | 2015-12-03 | Wageningen Universiteit | Targeted screening for novel disease resistance in plants |
US10301637B2 (en) | 2014-06-20 | 2019-05-28 | Cellectis | Potatoes with reduced granule-bound starch synthase |
US10988764B2 (en) | 2014-06-23 | 2021-04-27 | Monsanto Technology Llc | Compositions and methods for regulating gene expression via RNA interference |
WO2015200539A1 (en) | 2014-06-25 | 2015-12-30 | Monsanto Technology Llc | Methods and compositions for delivering nucleic acids to plant cells and regulating gene expression |
US10378012B2 (en) | 2014-07-29 | 2019-08-13 | Monsanto Technology Llc | Compositions and methods for controlling insect pests |
EP3174983A4 (en) | 2014-08-01 | 2017-12-13 | The New Zealand Institute for Plant and Food Research Limited | Methods and materials for producing coreless fruit |
BR102014023930A2 (en) | 2014-09-26 | 2016-05-17 | Ctc Ct De Tecnologia Canavieira S A | method for transforming a plant cell or plant tissue using agrobacterium, transgenic plant, transgenic cell or transgenic tissue, culture medium, and using a method for transforming a plant cell or plant tissue |
MX367829B (en) | 2014-11-04 | 2019-09-09 | Agresearch Ltd | Methods for plant improvement. |
MX2017005731A (en) | 2014-11-04 | 2017-07-28 | Agresearch Ltd | Methods for monocot plant improvement. |
CA2974716A1 (en) | 2014-11-12 | 2016-05-19 | Nmc, Inc. | Transgenic plants with engineered redox sensitive modulation of photosynthetic antenna complex pigments and methods for making the same |
US20180169211A1 (en) | 2014-11-13 | 2018-06-21 | Evaxion Biotech Aps | Peptides derived from acinetobacter baumannii and their use in vaccination |
WO2016100804A1 (en) | 2014-12-19 | 2016-06-23 | AgBiome, Inc. | Methods and compositions for providing resistance to glufosinate |
EP3037432B1 (en) | 2014-12-22 | 2020-06-17 | Dow AgroSciences LLC | Nucampholin nucleic acid molecules to control coleopteran insect pests |
US10570182B2 (en) | 2014-12-30 | 2020-02-25 | Dow Agrosciences Llc | Modified Cry1Ca toxins useful for control of insect pests |
EP3244918A2 (en) | 2015-01-12 | 2017-11-22 | Evaxion Biotech ApS | Proteins and nucleic acids useful in vaccines targeting klebsiella pneumoniae |
CA2974101A1 (en) | 2015-01-22 | 2016-07-28 | Monsanto Technology Llc | Compositions and methods for controlling leptinotarsa |
EP3250675A1 (en) | 2015-01-28 | 2017-12-06 | SABIC Global Technologies B.V. | Methods and compositions for high-efficiency production of biofuel and/or biomass |
EP3253780A1 (en) | 2015-02-04 | 2017-12-13 | Basf Plant Science Company GmbH | Method of increasing resistance against soybean rust in transgenic plants by increasing the scopoletin content |
US11421229B2 (en) | 2015-02-20 | 2022-08-23 | Baylor College Of Medicine | p63 inactivation for the treatment of heart failure |
JP6557989B2 (en) | 2015-02-23 | 2019-08-14 | 住友ゴム工業株式会社 | Vector containing gene encoding specific promoter and specific protein, transformed plant introduced with the vector, and method for improving polyisoprenoid production by introducing the vector into the plant |
JP6719174B2 (en) | 2015-02-23 | 2020-07-08 | 住友ゴム工業株式会社 | A vector containing a specific promoter and a gene encoding a specific protein, a transformed plant into which the vector is introduced, and a method for improving the production amount of polyisoprenoid by introducing the vector into the plant |
JP6557990B2 (en) | 2015-02-23 | 2019-08-14 | 住友ゴム工業株式会社 | Vector containing gene encoding specific promoter and specific protein, transformed plant introduced with the vector, and method for improving polyisoprenoid production by introducing the vector into the plant |
US20160264991A1 (en) | 2015-03-13 | 2016-09-15 | Dow Agrosciences Llc | Rna polymerase i1 nucleic acid molecules to control insect pests |
EP3283505A1 (en) | 2015-04-13 | 2018-02-21 | Fraunhofer Gesellschaft zur Förderung der angewandten Forschung E.V. | Novel aflatoxin and fungal infection control methods |
EP3283632A1 (en) | 2015-04-15 | 2018-02-21 | Dow Agrosciences LLC | Plant promoter for transgene expression |
RU2017137599A (en) | 2015-04-15 | 2019-05-15 | ДАУ АГРОСАЙЕНСИЗ ЭлЭлСи | VEGETABLE PROMOTER FOR THE TRANSGEN EXPRESSION |
EP3603651B1 (en) | 2015-05-22 | 2021-07-21 | STCube & Co., Inc. | Screening methods for targets for cancer therapy |
BR102016012010A2 (en) | 2015-05-29 | 2020-03-24 | Dow Agrosciences Llc | NUCLEIC ACID, RIBONUCLEIC ACID (RNA) AND DOUBLE-FILAMENT RIBONUCLEIC ACID (DSRNA) MOLECULE, CELL, PLANT AND SEED USES, PRIMARY PRODUCT, AS WELL AS METHODS TO CONTROL A POPULATION OF HOLIDAYS, OR HOSPITALS, OR HOSPITALS, OR HOSPITALS, OR HOSPITALS THE INCOME OF A CULTURE, AND TO PRODUCE A TRANSGENIC VEGETABLE CELL AND A TRANSGENIC PLANT |
CN107750125A (en) | 2015-06-02 | 2018-03-02 | 孟山都技术有限公司 | For by the composition and method in delivery of polynucleotides to plant |
WO2016196782A1 (en) | 2015-06-03 | 2016-12-08 | Monsanto Technology Llc | Methods and compositions for introducing nucleic acids into plants |
EP3317295B1 (en) | 2015-07-04 | 2022-05-18 | Evaxion Biotech A/S | Proteins and nucleic acids useful in vaccines targeting pseudomonas aeruginosa |
CN108026149B (en) | 2015-08-17 | 2022-04-29 | 美国陶氏益农公司 | Engineered CRY6A insecticidal proteins |
AU2016318051B2 (en) | 2015-09-04 | 2022-11-03 | Keygene N.V. | Diplospory gene |
IL241462A0 (en) | 2015-09-10 | 2015-11-30 | Yeda Res & Dev | Heterologous engineering of betalain pigments in plants |
US10837024B2 (en) | 2015-09-17 | 2020-11-17 | Cellectis | Modifying messenger RNA stability in plant transformations |
TW201718862A (en) | 2015-09-22 | 2017-06-01 | Dow Agrosciences Llc | Plant promoter and 3' UTR for transgene expression |
TW201718861A (en) | 2015-09-22 | 2017-06-01 | 道禮責任有限公司 | Plant promoter and 3'UTR for transgene expression |
WO2017059341A1 (en) | 2015-10-02 | 2017-04-06 | Monsanto Technology Llc | Recombinant maize b chromosome sequence and uses thereof |
AU2016342183B2 (en) | 2015-10-20 | 2022-03-03 | FUJIFILM Cellular Dynamics, Inc. | Methods for directed differentiation of pluripotent stem cells to immune cells |
AU2016340893B2 (en) | 2015-10-22 | 2019-06-27 | Corteva Agriscience Llc | Plant promoter for transgene expression |
TWI812584B (en) | 2015-10-30 | 2023-08-21 | 美國加利福尼亞大學董事會 | Methods of generating t-cells from stem cells and immunotherapeutic methods using the t-cells |
ES2892972T3 (en) | 2015-11-02 | 2022-02-07 | Univ Texas | Methods of CD40 activation and immune checkpoint blockade |
WO2017078935A1 (en) | 2015-11-04 | 2017-05-11 | Dow Agrosciences Llc | Plant promoter for transgene expression |
BR112018009270A2 (en) | 2015-11-06 | 2018-11-06 | Pioneer Hi Bred Int | vector, method of transforming a plant, kit |
AU2016349632A1 (en) | 2015-11-07 | 2018-05-24 | Multivir Inc. | Compositions comprising tumor suppressor gene therapy and immune checkpoint blockade for the treatment of cancer |
EP3390641B1 (en) | 2015-12-16 | 2023-08-09 | The New Zealand Institute for Plant and Food Research Limited | Compositions and methods for manipulating the development of plants |
UY37108A (en) | 2016-02-02 | 2017-08-31 | Cellectis | MODIFICATION OF THE COMPOSITION OF SOYBEAN OILS THROUGH DIRECTED KNOCKOUT OF THE FAD3A / B / C GENES |
BR112018016164A2 (en) | 2016-02-11 | 2019-01-22 | Pioneer Hi Bred Int | method of selecting a first plant or soybean germplasm, plasma or soybean germplasm, and kit for selecting at least one soybean plant |
WO2017144523A1 (en) | 2016-02-22 | 2017-08-31 | Evaxion Biotech Aps | Proteins and nucleic acids useful in vaccines targeting staphylococcus aureus |
US20190177391A1 (en) | 2016-03-31 | 2019-06-13 | Baylor Research Institute | Angiopoietin-like protein 8 (angptl8) |
US10563216B2 (en) | 2016-04-18 | 2020-02-18 | Bloomsburg University of Pennsylvania | Compositions and methods of delivering molecules to plants |
WO2017184727A1 (en) | 2016-04-21 | 2017-10-26 | Bayer Cropscience Lp | Tal-effector mediated herbicide tolerance |
EP3054014A3 (en) | 2016-05-10 | 2016-11-23 | BASF Plant Science Company GmbH | Use of a fungicide on transgenic plants |
CN109415737A (en) | 2016-05-25 | 2019-03-01 | 嘉吉公司 | For generating the engineered nucleic acid enzyme of mutation in plant |
CN116287385A (en) | 2016-06-16 | 2023-06-23 | 纽希得营养澳大利亚私人有限公司 | Inbred transgenic rape line NS-B50027-4 and seeds thereof |
EP3472281A4 (en) | 2016-06-16 | 2020-02-19 | Nuseed Pty Ltd | Elite event canola ns-b50027-4 |
WO2017216384A1 (en) | 2016-06-17 | 2017-12-21 | Evaxion Biotech Aps | Vaccination targeting ichthyophthirius multifiliis |
WO2017220787A1 (en) | 2016-06-24 | 2017-12-28 | Evaxion Biotech Aps | Vaccines against aearomonas salmonicida infection |
CN109642235B (en) | 2016-06-28 | 2023-12-29 | 孟山都技术公司 | Methods and compositions for genome modification in plants |
AU2017290805B2 (en) | 2016-07-01 | 2023-11-16 | Research Development Foundation | Elimination of proliferating cells from stem cell-derived grafts |
EP3487872A1 (en) | 2016-07-22 | 2019-05-29 | Evaxion Biotech ApS | Chimeric proteins for inducing immunity towards infection with s. aureus |
CN117604021A (en) | 2016-08-17 | 2024-02-27 | 孟山都技术公司 | Methods and compositions for dwarf plants for increasing harvestable yield by manipulating gibberellin metabolism |
EP3504334B1 (en) | 2016-08-26 | 2023-12-06 | Board of Trustees of Michigan State University | Transcription factors to improve resistance to environmental stress in plants |
IL247752A0 (en) | 2016-09-11 | 2016-11-30 | Yeda Res & Dev | Compositions and methods for regulating gene expression for targeted mutagenesis |
US20190225974A1 (en) | 2016-09-23 | 2019-07-25 | BASF Agricultural Solutions Seed US LLC | Targeted genome optimization in plants |
WO2018060881A1 (en) | 2016-09-27 | 2018-04-05 | University Of Florida Research Foundation, Inc. | Insect toxin delivery mediated by a densovirus coat protein |
EP3518657B1 (en) | 2016-10-03 | 2022-07-13 | Corteva Agriscience LLC | Plant promoter for transgene expression |
EP3519574B1 (en) | 2016-10-03 | 2022-01-19 | Corteva Agriscience LLC | Plant promoter for transgene expression |
WO2018067826A1 (en) | 2016-10-05 | 2018-04-12 | Cellular Dynamics International, Inc. | Generating mature lineages from induced pluripotent stem cells with mecp2 disruption |
BR112019006910A2 (en) | 2016-10-05 | 2019-07-02 | Fujifilm Cellular Dynamics Inc | methods for directed differentiation of pluripotent stem cells to hoz homozygous immune cells |
US10584350B2 (en) | 2016-10-27 | 2020-03-10 | Board Of Trustees Of Michigan State University | Structurally modified COI1 |
WO2018087301A1 (en) | 2016-11-10 | 2018-05-17 | Keygene N.V. | Methods for improving drought resistance in plants - kgdr06, kgdr26, kgdr25, kgdr42 and kgdr37 |
US11312972B2 (en) | 2016-11-16 | 2022-04-26 | Cellectis | Methods for altering amino acid content in plants through frameshift mutations |
WO2018101824A1 (en) | 2016-11-30 | 2018-06-07 | Universiteit Van Amsterdam | Plants comprising pathogen effector constructs |
CN110381997A (en) | 2016-12-12 | 2019-10-25 | 茂体外尔公司 | For treating and preventing the method and composition comprising gene-virus therapy and immunologic test point inhibitor of cancer and infectious diseases |
EP3342780A1 (en) | 2016-12-30 | 2018-07-04 | Dow AgroSciences LLC | Pre-mrna processing factor 8 (prp8) nucleic acid molecules to control insect pests |
EP3565576A1 (en) | 2017-01-05 | 2019-11-13 | Evaxion Biotech ApS | Vaccines targeting pseudomonas aeruginosa |
WO2018146323A1 (en) | 2017-02-13 | 2018-08-16 | Keygene N.V. | Method for altering ripening characteristics of fruit |
WO2018146322A1 (en) | 2017-02-13 | 2018-08-16 | Keygene N.V. | Method for altering ripening characteristics of fruit |
BR112019018175A2 (en) | 2017-03-07 | 2020-04-07 | BASF Agricultural Solutions Seed US LLC | molecule, cell, plant, seed, recombinant polypeptides, method for producing a polypeptide, plant, method for controlling herbs, use of nucleic acid and utility product |
BR112019018059A2 (en) | 2017-03-07 | 2020-08-04 | BASF Agricultural Solutions Seed US LLC | recombinant nucleic acid molecule, host cell, plants, transgenic seeds, recombinant polypeptide, method for producing a polypeptide, weed control method, use of nucleic acid and utility product |
US10883089B2 (en) | 2017-04-04 | 2021-01-05 | Wisconsin Alumni Research Foundation | Feruloyl-CoA:monolignol transferases |
US10883090B2 (en) | 2017-04-18 | 2021-01-05 | Wisconsin Alumni Research Foundation | P-coumaroyl-CoA:monolignol transferases |
KR20190141190A (en) | 2017-04-18 | 2019-12-23 | 후지필름 셀룰러 다이내믹스, 인코포레이티드 | Antigen-specific immune effector cells |
AU2018260469A1 (en) | 2017-04-25 | 2019-11-14 | Cellectis | Alfalfa with reduced lignin composition |
EP3630973A1 (en) | 2017-05-31 | 2020-04-08 | Tropic Biosciences UK Limited | Methods of selecting cells comprising genome editing events |
GB201708662D0 (en) | 2017-05-31 | 2017-07-12 | Tropic Biosciences Uk Ltd | Compositions and methods for increasing shelf-life of banana |
GB201708665D0 (en) | 2017-05-31 | 2017-07-12 | Tropic Biosciences Uk Ltd | Compositions and methods for increasing extractability of solids from coffee beans |
CA3073662A1 (en) | 2017-08-22 | 2019-02-28 | Napigen, Inc. | Organelle genome modification using polynucleotide guided endonuclease |
EP3676383A4 (en) | 2017-08-31 | 2021-09-08 | Dow Agrosciences LLC | Compositions and methods for expressing transgenes using regulatory elements from chlorophyll binding ab genes |
EP3684914A4 (en) | 2017-09-18 | 2021-06-02 | Futuragene Israel Ltd. | Tissue-specific expression control of della polypeptides |
DK3684930T3 (en) | 2017-09-19 | 2022-06-07 | Tropic Biosciences Uk Ltd | Modification of the specificity of non-coding plant RNA molecules for inactivation of gene expression |
WO2019083808A1 (en) | 2017-10-24 | 2019-05-02 | Basf Se | Improvement of herbicide tolerance to hppd inhibitors by down-regulation of putative 4-hydroxyphenylpyruvate reductases in soybean |
BR112020008092A2 (en) | 2017-10-24 | 2020-09-15 | BASF Agricultural Solutions Seed US LLC | method for checking tolerance to a GM herbicide and soy plant |
US11578334B2 (en) | 2017-10-25 | 2023-02-14 | Monsanto Technology Llc | Targeted endonuclease activity of the RNA-guided endonuclease CasX in eukaryotes |
EP3704140A1 (en) | 2017-10-31 | 2020-09-09 | Vilmorin & Cie | Wheat comprising male fertility restorer alleles |
WO2019086603A1 (en) | 2017-11-03 | 2019-05-09 | Interna Technologies B.V. | Mirna molecule, equivalent, antagomir, or source thereof for treating and/or diagnosing a condition and/or a disease associated with neuronal deficiency or for neuronal (re)generation |
US11612618B2 (en) | 2017-11-14 | 2023-03-28 | Henry Ford Health System | Compositions for use in the treatment and prevention of cardiovascular disorders resulting from cerebrovascular injury |
JP2021509815A (en) | 2018-01-05 | 2021-04-08 | オタワ ホスピタル リサーチ インスティチュート | Modified vaccinia vector |
WO2019145399A1 (en) | 2018-01-24 | 2019-08-01 | Evaxion Biotech Aps | Vaccines for prophylaxis of s. aureus infections |
US11905518B2 (en) | 2018-02-12 | 2024-02-20 | Curators Of The University Of Missouri | Small auxin upregulated (SAUR) gene for the improvement of root system architecture, waterlogging tolerance, drought resistance and yield in plants and methods of uses |
CN111836895A (en) | 2018-02-15 | 2020-10-27 | 孟山都技术公司 | Compositions and methods for increasing crop yield through trait stacking |
EP3752618A4 (en) | 2018-02-15 | 2021-11-03 | Monsanto Technology LLC | Compositions and methods for improving crop yields through trait stacking |
KR20200135986A (en) | 2018-03-19 | 2020-12-04 | 멀티비르 인코포레이티드 | Methods and compositions for cancer treatment comprising tumor suppressor gene therapy and CD122/CD132 agonists |
AU2019246415A1 (en) | 2018-03-30 | 2020-09-17 | Keygene N.V. | QTLs for powdery mildew resistance in melon |
JP2021519587A (en) | 2018-03-30 | 2021-08-12 | ユニバーシティ オブ ジュネーブ | MicroRNA expression constructs and their use |
BR112020020340A2 (en) | 2018-04-05 | 2021-01-05 | Keygene N.V. | IMPROVEMENT OF BROKER REGENERATION BY OVEREXPRESSION OF CHK GENES |
GB201805865D0 (en) | 2018-04-09 | 2018-05-23 | Innes John Centre | Genes |
NL2020823B1 (en) | 2018-04-25 | 2019-11-05 | Univ Delft Tech | NAD+ dependent 7ß-hydroxysteroid dehydrogenase |
GB201807192D0 (en) | 2018-05-01 | 2018-06-13 | Tropic Biosciences Uk Ltd | Compositions and methods for reducing caffeine content in coffee beans |
BR112020022160A2 (en) | 2018-05-01 | 2021-02-02 | Keygene N.V. | genes for hormone-free plant regeneration |
WO2019227030A1 (en) | 2018-05-24 | 2019-11-28 | Monsanto Technology Llc | Genome editing in plants |
EP3802839A1 (en) | 2018-06-07 | 2021-04-14 | The State of Israel, Ministry of Agriculture & Rural Development, Agricultural Research Organization (ARO) (Volcani Center) | Nucleic acid constructs and methods of using same |
CA3102975A1 (en) | 2018-06-07 | 2019-12-12 | The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center) | Methods of regenerating and transforming cannabis |
US20210238612A1 (en) | 2018-07-12 | 2021-08-05 | Keygene N.V. | Type v crispr/nuclease-system for genome editing in plant cells |
US10941428B2 (en) | 2018-09-12 | 2021-03-09 | Universidad Politécnica de Madrid | Reagents and methods for the expression of an active NifB protein and uses thereof |
US20220000932A1 (en) | 2018-09-28 | 2022-01-06 | Henry Ford Health System | Use of extracellular vesicles in combination with tissue plasminogen activator and/or thrombectomy to treat stroke |
EP3870207A1 (en) | 2018-10-22 | 2021-09-01 | Evaxion Biotech ApS | Vaccines targeting m. catharrhalis |
EP3874048A1 (en) | 2018-11-01 | 2021-09-08 | Keygene N.V. | Dual guide rna for crispr/cas genome editing in plants cells |
US11946057B2 (en) | 2018-12-20 | 2024-04-02 | Benson Hill, Inc. | Pre-conditioning treatments to improve plant transformation |
CA3123890A1 (en) | 2019-01-04 | 2020-07-09 | Cargill Incorporated | Engineered nucleases to generate mutations in plants |
WO2020169830A1 (en) | 2019-02-21 | 2020-08-27 | Keygene N.V. | Genotyping of polyploids |
US20220143168A1 (en) | 2019-02-27 | 2022-05-12 | Evaxion Biotech A/S | Vaccines targeting H. influenzae |
GB2595606B (en) | 2019-03-07 | 2022-09-21 | Univ California | CRISPR-Cas effector polypeptides and methods of use thereof |
GB201903521D0 (en) | 2019-03-14 | 2019-05-01 | Tropic Biosciences Uk Ltd | No title |
GB201903519D0 (en) | 2019-03-14 | 2019-05-01 | Tropic Biosciences Uk Ltd | Introducing silencing activity to dysfunctional rna molecules and modifying their specificity against a gene of interest |
GB201903520D0 (en) | 2019-03-14 | 2019-05-01 | Tropic Biosciences Uk Ltd | Modifying the specificity of non-coding rna molecules for silencing genes in eukaryotic cells |
NL2022905B1 (en) | 2019-04-09 | 2020-10-20 | Univ Delft Tech | Yeast with engineered Molybdenum co-factor biosynthesis |
US20220267386A1 (en) | 2019-05-07 | 2022-08-25 | Agventure B.V. | Self-compatibility in cultivated potato |
NL2023169B1 (en) | 2019-05-21 | 2020-12-01 | Univ Delft Tech | Biotin prototrophy |
US11447790B2 (en) | 2019-05-29 | 2022-09-20 | Altria Client Services Llc | Compositions and methods for producing tobacco plants and products having reduced or eliminated suckers |
WO2020239984A1 (en) | 2019-05-29 | 2020-12-03 | Keygene N.V. | Gene for parthenogenesis |
JP2020195326A (en) | 2019-06-03 | 2020-12-10 | 住友ゴム工業株式会社 | Method of producing natural rubber, transgenic plant, method of producing pneumatic tire, and method of producing rubber product |
US20230032478A1 (en) | 2019-07-11 | 2023-02-02 | The Regents Of The University Of California | Methods for Improved Regeneration of Plants Using Growth-Regulating Factor (GRF), GRF-Interacting Factor (GIF), or Chimeric GRF-GIF |
JP6923166B2 (en) | 2019-07-16 | 2021-08-18 | 住友ゴム工業株式会社 | Fusion protein, substance manufacturing method, vector, transformed cell, pneumatic tire manufacturing method and rubber product manufacturing method |
WO2021015616A1 (en) | 2019-07-22 | 2021-01-28 | Wageningen Universiteit | Lrr-rlkii receptor kinase interaction domains |
CN114745946B (en) | 2019-10-10 | 2024-03-08 | 奥驰亚客户服务有限公司 | Pale yellow locus and application thereof in tobacco |
US11939588B2 (en) | 2019-10-17 | 2024-03-26 | Board Of Trustees Of Michigan State University | Elevated resistance to insects and plant pathogens without compromising seed production |
EP4045044A1 (en) | 2019-10-18 | 2022-08-24 | The Regents Of The University Of California | Plxdc activators and their use in the treatment of blood vessel disorders |
JP2021080204A (en) | 2019-11-19 | 2021-05-27 | 住友ゴム工業株式会社 | Fusion proteins, methods for producing substance, vectors, transformed cells, methods for manufacturing pneumatic tires, and methods for manufacturing rubber products |
EP3825408A1 (en) | 2019-11-19 | 2021-05-26 | FRAUNHOFER-GESELLSCHAFT zur Förderung der angewandten Forschung e.V. | Methods of multi-species insect pest control |
US11326176B2 (en) | 2019-11-22 | 2022-05-10 | Mozza Foods, Inc. | Recombinant micelle and method of in vivo assembly |
WO2021113644A1 (en) | 2019-12-05 | 2021-06-10 | Multivir Inc. | Combinations comprising a cd8+ t cell enhancer, an immune checkpoint inhibitor and radiotherapy for targeted and abscopal effects for the treatment of cancer |
EP4075951A1 (en) | 2019-12-19 | 2022-10-26 | Keygene N.V. | Germacrene a synthase mutants |
EP4087593A1 (en) | 2020-01-06 | 2022-11-16 | Evaxion Biotech A/S | Vaccines targeting neisseria gonorrhoeae |
US20230107598A1 (en) | 2020-02-28 | 2023-04-06 | Pioneer Hi-Bred International, Inc. | Sorghum doubled haploid production system |
WO2021177074A1 (en) | 2020-03-05 | 2021-09-10 | 住友ゴム工業株式会社 | Method for producing polyisoprenoid, vector, transformed plant, method for producing pneumatic tire, and method for producing rubber product |
EP4157864A1 (en) | 2020-05-27 | 2023-04-05 | Antion Biosciences SA | Adapter molecules to re-direct car t cells to an antigen of interest |
BR112022024031A2 (en) | 2020-05-29 | 2023-01-31 | Fujifilm Cellular Dynamics Inc | RETINAL PIGMENTED EPITHELIUM AND PHOTORECEPTOR DUAL CELL AGGREGATES, AND METHODS OF USE THEREOF |
IL298254A (en) | 2020-05-29 | 2023-01-01 | Fujifilm Cellular Dynamics Inc | Bilayer of retinal pigmented epithelium and photoreceptors and use thereof |
WO2021260632A1 (en) | 2020-06-24 | 2021-12-30 | Benson Hill, Inc. | Plant cell treatments to improve plant transformation |
WO2022047022A1 (en) | 2020-08-26 | 2022-03-03 | Vindara, Inc. | METHOD and/or COMPOSITIONS FOR LETTUCE (LACTUCA SATIVA) BREEDING and/or VARIETIES DEVELOPED THEREBY |
JP2023544753A (en) | 2020-10-13 | 2023-10-25 | キージーン ナムローゼ フェンノートシャップ | Modified promoters of parthenogenetic genes |
WO2022093977A1 (en) | 2020-10-30 | 2022-05-05 | Fortiphyte, Inc. | Pathogen resistance in plants |
US20220162623A1 (en) | 2020-11-20 | 2022-05-26 | AgBiome, Inc. | Compositions and methods for incorporation of dna into the genome of an organism |
US20220243215A1 (en) | 2021-02-03 | 2022-08-04 | Altria Client Services Llc | Terpene production in plants |
EP4291216A1 (en) | 2021-02-09 | 2023-12-20 | University of Houston System | Oncolytic virus for systemic delivery and enhanced anti-tumor activities |
CA3207574A1 (en) | 2021-02-11 | 2022-08-18 | Wouter Leonard De Laat | Curing disease by transcription regulatory gene editing |
EP4294445A1 (en) | 2021-02-19 | 2023-12-27 | Pfizer Inc. | Methods of protecting rna |
GB202103256D0 (en) | 2021-03-09 | 2021-04-21 | Tropic Biosciences Uk Ltd | Method for silencing genes |
WO2022219175A1 (en) | 2021-04-15 | 2022-10-20 | Keygene N.V. | Mobile endonucleases for heritable mutations |
CA3212047A1 (en) | 2021-04-15 | 2022-10-20 | Jeroen Stuurman | Co-regeneration recalcitrant plants |
CA3216719A1 (en) | 2021-05-03 | 2022-11-10 | Astellas Institute For Regenerative Medicine | Methods of generating mature corneal endothelial cells |
CN117716020A (en) | 2021-05-07 | 2024-03-15 | 安斯泰来再生医药协会 | Method for producing mature hepatocytes |
EP4347796A1 (en) | 2021-05-26 | 2024-04-10 | Fujifilm Cellular Dynamics, Inc. | Methods to prevent rapid silencing of genes in pluripotent stem cells |
US20220386549A1 (en) | 2021-06-02 | 2022-12-08 | Nutrien Ag Solutions, Inc. | Inbred rice line dg263l |
US20220387581A1 (en) | 2021-06-03 | 2022-12-08 | Mazen Animal Health Inc. | Oral administration of coronavirus spike protein for altering cytokine levels and providing passive immunity to newborn pigs |
WO2023275255A1 (en) | 2021-07-02 | 2023-01-05 | Tropic Biosciences UK Limited | Delay or prevention of browning in banana fruit |
GB202109586D0 (en) | 2021-07-02 | 2021-08-18 | Tropic Biosciences Uk Ltd | Method for editing banana genes |
AU2022307747A1 (en) | 2021-07-05 | 2024-01-25 | Evaxion Biotech A/S | Vaccines targeting neisseria gonorrhoeae |
GB202112865D0 (en) | 2021-09-09 | 2021-10-27 | Tropic Biosciences Uk Ltd | Resistance to black sigatoka disease in banana |
GB202112866D0 (en) | 2021-09-09 | 2021-10-27 | Tropic Biosciences Uk Ltd | Resistance to fusarium wilt in a banana |
JP2023040705A (en) | 2021-09-10 | 2023-03-23 | 住友ゴム工業株式会社 | Method for producing trans-polyisoprenoid, vector, transgenic organism, method for producing pneumatic tire, and method for producing rubber product |
WO2023089556A1 (en) | 2021-11-22 | 2023-05-25 | Pfizer Inc. | Reducing risk of antigen mimicry in immunogenic medicaments |
EP4186917A1 (en) | 2021-11-29 | 2023-05-31 | Keygene N.V. | Tobamovirus resistant plants |
WO2023111225A1 (en) | 2021-12-17 | 2023-06-22 | Keygene N.V. | Double decapitation of plants |
GB2614309A (en) | 2021-12-24 | 2023-07-05 | Stratosvir Ltd | Improved vaccinia virus vectors |
WO2023144779A1 (en) | 2022-01-28 | 2023-08-03 | Pfizer Inc. | Coronavirus antigen variants |
JP2023127870A (en) | 2022-03-02 | 2023-09-14 | 住友ゴム工業株式会社 | Mutant cis-prenyltransferase (cpt) family protein, method for producing polyisoprenoid, vector, transgenic plant, method for producing pneumatic tire, and method for producing rubber product |
JP2023127868A (en) | 2022-03-02 | 2023-09-14 | 住友ゴム工業株式会社 | Mutant cis-prenyltransferase (cpt) family protein, method for producing polyisoprenoid, vector, transgenic plant, method for producing pneumatic tire, and method for producing rubber product |
JP2023127869A (en) | 2022-03-02 | 2023-09-14 | 住友ゴム工業株式会社 | Mutant cis-prenyltransferase (cpt) family protein, method for producing polyisoprenoid, vector, transgenic plant, method for producing pneumatic tire, and method for producing rubber product |
US20230295661A1 (en) | 2022-03-16 | 2023-09-21 | University Of Houston System | Persistent hsv gene delivery system |
US20230392160A1 (en) | 2022-04-12 | 2023-12-07 | John Innes Centre | Compositions and methods for increasing genome editing efficiency |
GB202206507D0 (en) | 2022-05-04 | 2022-06-15 | Antion Biosciences Sa | Expression construct |
WO2023213393A1 (en) | 2022-05-04 | 2023-11-09 | Evaxion Biotech A/S | Staphylococcal protein variants and truncates |
WO2023230433A1 (en) | 2022-05-23 | 2023-11-30 | Altria Client Services Llc | Methods and compositions for regulating alkaloids in tobacco field |
US20240003871A1 (en) | 2022-06-29 | 2024-01-04 | FUJIFILM Cellular Dynamics, Inc. | Ipsc-derived astrocytes and methods of use thereof |
WO2024008763A2 (en) | 2022-07-05 | 2024-01-11 | Keygene N.V. | Orobanche resistant plants |
WO2024023067A1 (en) | 2022-07-25 | 2024-02-01 | Koninklijke Nederlandse Akademie Van Wetenschappen | Curing disease by transcription regulatory gene editing |
WO2024031038A1 (en) | 2022-08-05 | 2024-02-08 | Altria Client Services Llc | Methods and compositions for regulating alkaloids in tobacco |
WO2024047210A1 (en) | 2022-09-01 | 2024-03-07 | Keygene N.V. | Hydrogel-carriers for induction of plant morphogenesis |
US20240093220A1 (en) | 2022-09-09 | 2024-03-21 | Friedrich Alexander Universität Erlangen-Nürnberg | Plant regulatory elements and uses thereof |
GB202305021D0 (en) | 2023-04-04 | 2023-05-17 | Tropic Biosciences Uk Ltd | Methods for generating breaks in a genome |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1992009696A1 (en) * | 1990-11-23 | 1992-06-11 | Plant Genetic Systems, N.V. | Process for transforming monocotyledonous plants |
JPH04222527A (en) | 1990-12-19 | 1992-08-12 | Japan Tobacco Inc | Transformation of tomato |
WO1992014828A1 (en) * | 1991-02-14 | 1992-09-03 | Svalöf Ab | Method for genetic transformation of tissue organs from monocotyledonous plants |
EP0504869A2 (en) | 1991-03-20 | 1992-09-23 | Japan Tobacco Inc. | Tomato resistant to cucumber mosaic virus and method for transforming tomato |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0290799B9 (en) * | 1983-01-13 | 2004-09-01 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Transgenic dicotyledonous plant cells and plants |
NL8300698A (en) * | 1983-02-24 | 1984-09-17 | Univ Leiden | METHOD FOR BUILDING FOREIGN DNA INTO THE NAME OF DIABIC LOBAL PLANTS; AGROBACTERIUM TUMEFACIENS BACTERIA AND METHOD FOR PRODUCTION THEREOF; PLANTS AND PLANT CELLS WITH CHANGED GENETIC PROPERTIES; PROCESS FOR PREPARING CHEMICAL AND / OR PHARMACEUTICAL PRODUCTS. |
NL8401048A (en) * | 1984-04-03 | 1985-11-01 | Rijksuniversiteit Leiden En Pr | METHOD FOR BUILDING FOREIGN DNA INTO THE NAME OF LOBELIC PLANTS. |
NL8401780A (en) * | 1984-06-04 | 1986-01-02 | Rijksuniversiteit Leiden En Pr | METHOD FOR BUILDING FOREIGN DNA INTO THE GENOMES OF PLANTS |
US5149645A (en) * | 1984-06-04 | 1992-09-22 | Rijksuniversiteit Leiden | Process for introducing foreign DNA into the genome of plants |
US5177010A (en) * | 1986-06-30 | 1993-01-05 | University Of Toledo | Process for transforming corn and the products thereof |
CA1341455C (en) * | 1986-06-30 | 2004-04-27 | Stephen L. Goldman | Process for transforming gramineae and the products thereof |
US5187073A (en) * | 1986-06-30 | 1993-02-16 | The University Of Toledo | Process for transforming gramineae and the products thereof |
EP0419533A1 (en) * | 1988-06-01 | 1991-04-03 | THE TEXAS A&M UNIVERSITY SYSTEM | Method for transforming plants via the shoot apex |
-
1993
- 1993-07-06 DE DE69334225T patent/DE69334225D1/en not_active Expired - Lifetime
- 1993-07-06 EP EP93914958A patent/EP0604662B1/en not_active Expired - Lifetime
- 1993-07-06 AT AT93914958T patent/ATE398679T1/en not_active IP Right Cessation
- 1993-07-06 EP EP08009937A patent/EP1983056A1/en not_active Ceased
- 1993-07-06 US US08/193,058 patent/US5591616A/en not_active Expired - Lifetime
- 1993-07-06 DK DK93914958T patent/DK0604662T3/en active
- 1993-07-06 WO PCT/JP1993/000925 patent/WO1994000977A1/en active IP Right Grant
-
1994
- 1994-04-18 CA CA002121545A patent/CA2121545C/en not_active Expired - Lifetime
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1992009696A1 (en) * | 1990-11-23 | 1992-06-11 | Plant Genetic Systems, N.V. | Process for transforming monocotyledonous plants |
JPH04222527A (en) | 1990-12-19 | 1992-08-12 | Japan Tobacco Inc | Transformation of tomato |
WO1992014828A1 (en) * | 1991-02-14 | 1992-09-03 | Svalöf Ab | Method for genetic transformation of tissue organs from monocotyledonous plants |
EP0504869A2 (en) | 1991-03-20 | 1992-09-23 | Japan Tobacco Inc. | Tomato resistant to cucumber mosaic virus and method for transforming tomato |
Non-Patent Citations (60)
Title |
---|
BYTEBIER B. ET AL., PROC. NATL. ACAD. SCI. USA, vol. 84, 1987, pages 5345 - 5349 |
CABOCHE M., PHYSIOL. PLANT., vol. 79, 1990, pages 173 - 176 |
CHAN, M-T., ET AL.: "Agrobacterium-mediated production of transgenic rice plants expressing a chimeric alpha-amylase promoter/beta-glucuronidase gene", PLANT MOLECULAR BIOLOGY, vol. 22, no. 3, June 1993 (1993-06-01), pages 491 - 506, XP009006601 * |
CHAN, M-T., ET AL.: "Transformation of indica rice (Oryza sativa L.) mediated by Agrobacterium tumefaciens", PLANT CELL PHYSIOL., vol. 33, no. 5, 1 January 1992 (1992-01-01), pages 577 - 583, XP000562121 * |
CHRISTOU P. ET AL., BIO/TECHNOL, vol. 9, 1991, pages 957 - 962 |
CHU C.C.: "Proc. Symp. Plant Tissue Culture", 1978, article "Science Press Peking", pages: 43 - 50 |
CHU C.C.: "Proc. Symp. Plant Tissue Culture", 1987, SCIENCE PRESS PEKING, pages: 43 - 50 |
DATTA S.K. ET AL., BIO/TECHNOL., vol. 8, 1990, pages 736 - 740 |
DE CLEENE M, BOT. REV., vol. 42, 1976, pages 389 - 466 |
DE GREVE H.H. ET AL., PLASMID, vol. 6, 1981, pages 235 - 248 |
DITTA G. ET AL., PROC. NATL. ACAD. SCI. USA, vol. 77, 1980, pages 7347 - 7351 |
DRLICA K.A.; KADO C.I., PROC. NATL. ACAD. SCI. USA, vol. 71, 1974, pages 3677 - 3681 |
FROMM M.E. ET AL., BIO/TECHNOL, vol. 8, 1990, pages 833 - 839 |
GORDON-KAMM W.J. ET AL., PLANT CELL, vol. 2, 1990, pages 603 - 618 |
GOULD J ET AL: "TRANSFORMATION OF ZEA-MAYS L. USING AGROBACTERIUM-TUMEFACIENS AND THE SHOOT APEX", PLANT PHYSIOLOGY, AMERICAN SOCIETY OF PLANT PHYSIOLOGISTS, ROCKVILLE, MD, US, vol. 95, no. 2, 1 January 1991 (1991-01-01), pages 426 - 434, XP002179154, ISSN: 0032-0889 * |
GOULD J. ET AL., PLANT PHYSIOL., vol. 95, 1991, pages 426 - 434 |
GOULD J., PLANT PHYSIOL., vol. 95, 1991, pages 426 - 434 |
GRIMSLEY ET AL., BIO/TECHNOL., vol. 6, 1988, pages 185 - 189 |
GRIMSLEY ET AL., MOL. GEN. GENET., vol. 217, pages 309 - 316 |
GRIMSLEY ET AL., NATURE, vol. 325, 1987, pages 177 - 179 |
GRITZ L.; DAVIS J., GENE, vol. 25, 1983, pages 179 - 188 |
HERRERA-ESTERELLA L. ET AL., EMBO J., vol. 2, 1983, pages 987 - 995 |
HIEI YUKOH ET AL: "Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA", PLANT JOURNAL, BLACKWELL SCIENTIFIC PUBLICATIONS, OXFORD, GB, vol. 6, no. 2, 1 January 1994 (1994-01-01), pages 271 - 282, XP002147288, ISSN: 0960-7412 * |
HOOD E.E. ET AL., BIO/TECHNOL., vol. 2, 1984, pages 702 - 709 |
HOOD E.E. ET AL., J. BACTERIOL, vol. 168, 1986, pages 1283 - 1290 |
HORCH R. H. ET AL., SCIENCE, vol. 223, 1984, pages 496 - 498 |
JIN S. ET AL., J. BACTERIOL, vol. 169, 1987, pages 4417 - 4425 |
KINOSHITA, PLANT CELL TISSUE ORGAN CULT, vol. 12, 1988, pages 311 - 314 |
KOMARI ET AL., THEORETICAL AND APPLIED GENETICS, vol. 77, 1989, pages 547 - 552 |
KOMARI T. ET AL., J. BACTERIOL., vol. 166, 1986, pages 88 - 94 |
KOMARI T., PLANT SCIENCE, vol. 60, 1989, pages 223 - 229 |
KOMARI T: "TRANSFORMATION OF CULTURED CELLS OF CHENOPODIUM QUINOA BY BINARY VECTORS THAT CARRY A FRAGMENT OF DNA FROM THE VIRULENCE REGION OF PTIBO542", PLANT CELL REPORTS, SPRINGER VERLAG, DE, vol. 9, 1 January 1990 (1990-01-01), pages 303 - 306, XP000918219, ISSN: 0721-7714 * |
LEDOUX L. ET AL., NATURE, vol. 249, 1974, pages 17 - 21 |
LEE, N., ET AL.: "Efficient transformation and regeneration of rice small cell groups", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA, no. 88, 1991, WASHINGTON US, pages 6389 - 6393, XP000217486 * |
LI B-J, ET AL.: "STUDIES ON INTRODUCTION OF FOREIGN GENES INTO CULTURED CELLS OF ORYZA-SATIVA INDICA USING AGROBACTERIUM TI PLASMID SYSTEM.", SCI CHINA SER B CHEM LIFE SCI EARTH SCI 34 (1). 1991. 54-63., 1991, XP000560855 * |
LINSMAIER, E.; SKOOG, F., PHYSIOL. PLANT, vol. 18, 1965, pages 100 - 127 |
LUO; WU, PLANT MOL. BIOL. REP, vol. 6, 1988, pages 165 |
LUPOTTO, E.; LUSARDI, M.C., MAYDICA, vol. XXXIII, 1988, pages 163 - 177 |
MOONEY P.A. ET AL., PLANT CELL, TISSUE, ORGAN CULTURE, vol. 25, 1991, pages 209 - 218 |
MOONEY, P.A., ET AL.: "Agrobacterium tumefaciens-gene transfer into wheat tissues", PLANT CELL TISSUE AND ORGAN CULTURE, vol. 25, 1991, pages 209 - 218, XP000560763 * |
MURASHIGE, T.; SKOOG, F., PHYSIOL. PLANT., vol. 15, 1962, pages 473 - 497 |
MURASHIGE; SKOOG, PHYSIOL. PLANT, vol. 15, 1962, pages 473 - 497 |
NAKAMURA ET AL., PLANT BIOTECHNOLOGY II, 1991, pages 123 - 132 |
NEUHAUS G. ET AL., THEOR. APPL. GENET., vol. 75, 1987, pages 30 - 36 |
OHTA S. ET AL.: "Plant Cell Physiol.", vol. 31, 1990, DR. NAKAMURA OF NAGOYA UNIVERSITY, pages: 805 - 813 |
PIETRAZAK ET AL., NUCLEIC ACIDS RESEARCH, vol. 14, 1986, pages 5857 - 5868 |
POTRYKUS 1., BIO/TECHNOL., vol. 8, 1990, pages 535 - 543 |
POTRYKUS I, BIO/TECHNOL, vol. 8, 1990, pages 535 - . 543 |
RAINERI D M ET AL: "RESEARCH PAPERS/ AGROBACTERIUM-MEDIATED TRANSFORMATION OF RICE (ORYZA SATIVA L.)", BIOTECHNOLOGY, BUTTERWORTHS, LONDON, GB, vol. 8, 1 January 1990 (1990-01-01), pages 33 - 38, XP000918272, ISSN: 0740-7378 * |
RAINERI ET AL., BIO/TECHNOL., vol. 8, 1990, pages 33 - 38 |
RHODES C.A. ET AL., SCIENCE, vol. 240, 1989, pages 204 - 207 |
SAITO Y. ET AL., THEOR. APPL. GENET, vol. 83, 1992, pages 679 - 683 |
SAMBROOK ET AL.: "Molecular Cloning", 1989, COLD SPRING HARBOR LABORATORY PRESS |
SCHAFEW ET AL., NATURE, vol. 327, 1987, pages 529 - 532 |
SHIMAMOTO K. ET AL., NATURE, vol. 338, 1989, pages 274 - 276 |
TOPFER R. ET AL., PLANT CELL, vol. 1, 1989, pages 133 - 139 |
TORIYAMA K. ET AL., BIO/TECHNOL, vol. 6, 1988, pages 1072 - 1074 |
TORIYAMA; HINATA, PLANT SCIENCE, vol. 41, 1985, pages 179 - 183 |
XU Y: "INTERACTION AND TRANSFORMATION OF CEREAL CELLS WITH PHENOLICS-PRETREATED AGROBACTERIUM-TUMEFACIENS.", CHIN J BOT 2 (2). 1990. 81-87., 1990, XP000560825 * |
ZHANG W. ET AL., THEOR. APPL. GENET, vol. 76, 1988, pages 835 - 840 |
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EP0604662A4 (en) | 1996-04-17 |
ATE398679T1 (en) | 2008-07-15 |
US5591616A (en) | 1997-01-07 |
DK0604662T3 (en) | 2008-10-20 |
AU667939B2 (en) | 1996-04-18 |
DE69334225D1 (en) | 2008-07-31 |
WO1994000977A1 (en) | 1994-01-20 |
CA2121545A1 (en) | 1995-01-07 |
EP0604662B1 (en) | 2008-06-18 |
CA2121545C (en) | 2006-10-24 |
AU4513493A (en) | 1994-01-31 |
EP0604662A1 (en) | 1994-07-06 |
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